Bispecific hiv-1 neutralizing antibodies

ABSTRACT

In various embodiments, the present invention relates generally to using bispecific antibodies in the prevention and treatment of HIV.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority to and the benefit of, and incorporatesherein by reference in its entirety, U.S. Provisional Patent ApplicationNo. 61/910,685, entitled IMPROVED HIV-1-NEUTRALIZING ANTIBODY POTENCYAND BREADTH VIA CELL RECEPTOR ANCHORING USING BISPECIFIC ANTIBODIES WITHNATIVE ARCHITECTURE, which was filed on Dec. 2, 2013.

FIELD OF THE INVENTION

In various embodiments, the present invention relates generally to usingbispecific antibodies in the prevention and treatment of HIV.

BACKGROUND

Passive immunization with antibodies (Abs) is a recognized method ofprophylaxis and treatment of infectious diseases. This approach mayinvolve preparing human immunoglobulins from donors who recovered froman infectious disease and utilizing such preparations, containing Absspecific for the infectious organism, to protect a recipient against thesame disease. Alternatively, therapeutic antibodies can be made byimmunizing mice with an antigen, and then engineering/humanizing themouse Ab into a human version. Monoclonal antibodies (mAbs) arehomogeneous in terms of physical characteristics and immunochemicalreactivity, and so offer the possibility of absolute specific activity.

That specificity can ultimately be a limitation for some targets, sopractitioners have developed “bispecific” mAbs composed of fragments oftwo different mAbs and which bind to two different types of antigen.This facilitates binding to antigens expressed only weakly, for example.Some bispecific mAbs can stimulate strong immune responses, limitingtheir clinical application. One recent approach to ameliorating thiseffect is “CrossMab” methodology, a bispecific antibody format thatadopts a more native antibody-like structure.

The prospects for generating a highly potent bispecific or bivalentantibody against a pathogen, such as HIV, for clinical use involves manyuncertainties. The low spike density and spike structure on HIV mayimpede bivalent binding of antibodies to HIV, for example, and thegeometry and spatial relationship of cell surface anchoring are notwell-characterized. Nor is it known whether sufficient epitopeaccessibility on the HIV envelope exists. CrossMab bispecific antibodiesthat are anchored to a host cell membrane offer the possibility ofimproved local antibody concentration, targeting of sequential and/orinterdependent entry steps, and compensating for monovalent binding.

SUMMARY

In one aspect, the present invention pertains to a bispecific antibodyfor neutralizing HIV. The bispecific antibody includes portions of afirst and a second antibody, in which the first antibody binds to a HIVenvelope protein. In certain embodiments, the first antibody is selectedfrom PGT145, PG9, PGT128, PGT121, 10-1074, 3BNC117, VRC01, PGT151, 4E10,10E8 and a variant thereof. In certain embodiments, the bispecificantibody includes portions of a second antibody, in which the secondantibody binds to a cell membrane protein. For example, the secondantibody may binds to a cell receptor protein or a cell membraneco-receptor protein. In an embodiment, the second antibody is selectedfrom a CD4 antibody, a CCR5 antibody and a CXCR4 antibody, such as Pro140, ibalizumab, 515H7, or a variant thereof. In various embodiments,the bispecific antibody has a CrossMab format.

In another aspect, the present invention provides a bispecific antibodyincluding portions of a first antibody and a second antibody, whereinthe first antibody binds to a HIV envelope protein and the secondantibody binds to a cell membrane protein. In various embodiments, thebispecific antibody has a CrossMab format.

In various embodiments, pharmaceutical compositions including thebispecific antibodies disclosed herein are also provided. Thepharmaceutical composition may be formulated for oral, intranasal,pulmonary, intradermal, transdermal, subcutaneous, intramuscular,intraperitoneal, or intravenous delivery.

In a further aspect, methods for neutralizing HIV are provided. Themethods include the steps of contacting an antigen binding site with abispecific antibody that binds a HIV envelope protein and contactinganother antigen binding site with a bispecific antibody that binds acell membrane protein.

In another aspect, methods for treating a patient infected with HIV arealso provided. The methods include administering to the patient any ofthe bispecific antibodies or pharmaceutical compositions as disclosedherein. In an embodiment, the patient is human.

BRIEF DESCRIPTION OF THE DRAWINGS

In the drawings, like reference characters generally refer to the sameparts throughout the different views. Also, the drawings are notnecessarily to scale, with an emphasis instead generally being placedupon illustrating the principles of the invention. In the followingdescription, various embodiments of the present invention are describedwith reference to the following drawings, in which:

FIG. 1 is a diagram illustrating a CrossMab antibody derived from twoIgG monoclonal antibodies.

FIG. 2A is a diagram illustrating an iMab antibody (shorthand for themonoclonal antibody ibalizumab) that targets CD4 and a Pro 140 antibodythat targets CCR5.

FIG. 2B is a diagram illustrating mAbs that target the HIV envelopegp120.

FIG. 3 is a graph comparing the maximum percentage inhibition (MPI)against cell-to-cell HIV transmission using a combination of iMab and10E8 antibodies with CrossMab bispecific 10E8/iMab antibodies. Exceptotherwise stated, all iMab-based bispecific antibodies were constructedusing the MV1 variant.

FIG. 4 is a series of graphs comparing the inhibition of various strainsof X4 and dual-tropic HIV using varying concentrations of 10E8, Pro 140or 10E8/P140 antibodies. P140 is shorthand for Pro 140.

FIG. 5 is a series of graphs comparing the inhibition of various strainsof HIV using varying concentrations of 10E8, Pro 140, 10E8/P140 or acombination of the individual 10E8 and Pro 140 monoclonal antibodies.

FIG. 6 is a series of graphs comparing the inhibition of various strainsof HIV using varying concentrations of 10E8, X19, 10E8/X19 or 10E8/P140antibodies.

FIG. 7 is a series of graphs comparing the inhibition of various strainsof HIV using varying concentrations of 10E8, Pro 140, 10E8/P140 and10E8/αHer2 antibodies.

FIG. 8A is a graph comparing the binding of CrossMab bispecificantibodies 10E8/iMab and Δ10E8/iMab to the HIV-1 glycoprotein MPER.

FIG. 8B is a series of graphs comparing the inhibition percentages of10E8 (light gray lines) and Δ10E8 (dark gray lines) against iMabresistant R5 viruses (panel A) and X4 viruses (panel B), as well as theinhibition percentages of 10E8/iMab (light gray lines) and Δ10E8/iMab(dark gray lines) against iMab resistant R5 viruses (panel C) and X4viruses (panel D).

FIG. 9 is a series of graphs comparing the inhibition of various strainsof HIV using varying concentrations of 10E8, Δ10E8, 4E10, 10E8/P140,Δ10E8/P140 and 4E10/P140 antibodies.

FIG. 10 is a graph comparing the antiviral coverage of the CrossMabantibodies 10E8/Pro140 and 10E8/iMab, their parental monoclonalantibodies 10E8, Pro140 and iMab, and various other HIVenvelope-targeting monoclonal antibodies against a large panel of HIVenvelope pseudotyped viruses.

FIG. 11 is a series of graphs comparing the maximum percentageinhibition (MPI) of a large panel of HIV envelope pseudotyped viruseswith the monoclonal antibody iMab (grey bars in all panels) and theCrossMab antibodies PGT145/ibalizumab (145/iMab; top left panel),PGT128/ibalizumab (128/iMab; top center panel), PGT151/ibalizumab(151/iMab; top right panel), 3BNC117/ibalizumab (117/iMab; bottom leftpanel) and 10E8/ibalizumab (10E8/iMab; bottom right panel).

FIG. 12 is a series of graphs comparing the maximum percentageinhibition (MPI) and IC80 antibody concentrations of the CrossMabantibodies PGT145/ibalizumab (145/iMab; top left panel),PGT128/ibalizumab (128/iMab; top center panel), PGT151/ibalizumab(151/iMab; top right panel), 3BNC117/ibalizumab (117/iMab; bottom leftpanel) and 10E8/ibalizumab (10E8/iMab; bottom right panel) against alarge panel of HIV envelope pseudotyped viruses.

FIG. 13 is a series of graphs comparing the IC80 antibody concentrationsfor iMab-and Pro140-based CrossMab bispecific antibodies and theirparent antibodies for PGT145/iMab and PGT145/Pro140 (top left panel),3BNC117/iMab and 3BNC117/Pro140 (top center panel), PGT128/iMab andPGT128/Pro140 (top right panel), PGT151/iMab and PGT151/Pro140 (bottomleft panel) and 10E8/iMab and 10E8/Pro140 (bottom right panel).

FIG. 14 is a series of graphs comparing the IC50 antibody concentrationsfor iMab-and Pro140-based CrossMab bispecific antibodies and theirparent antibodies for PGT145/iMab and PGT145/Pro140 (top left panel),3BNC117/iMab and 3BNC117/Pro140 (top center panel), PGT128/iMab andPGT128/Pro140 (top right panel), PGT151/iMab and PGT151/Pro140 (bottomleft panel) and 10E8/iMab and 10E8/Pro140 (bottom right panel).

FIG. 15 is a graph displaying the IC80 antibody concentrations foriMab-based CrossMab bispecific antibodies and their parent antibodiesagainst cell-to-cell transmission of HIV for 10E8/iMab (top left panel),3BNC117/iMab (top center panel), PGT145/iMab (top right panel),PGT128/iMab (bottom left panel) and PGT151/iMab (bottom right panel).

FIG. 16 is a graph displaying the maximum percent inhibition (MPI) ofCrossMab bispecific antibodies and parental antibodies againstcell-to-cell transmission of HIV.

FIG. 17, left panel, is a graph comparing the inhibition of an HIVstrain against varying concentrations of 10E8, Pro 140, 10E8/P140CrossMab bispecific antibody, and a combination of individual 10E8 andPro 140 monoclonal antibodies. FIG. 17, right panel, is a graphcomparing the inhibition of an HIV strain against varying concentrationsof iMab, 10E8, 10E8/iMab CrossMab bispecific antibody, and a combinationof individual 10E8 and iMab monoclonal antibodies.

FIG. 18, top panel, is a series of graphs comparing the inhibition ofvarious HIV R5 strains against varying concentrations of 10E8, Pro140,10E8/P140 and 10E8/515H7 antibodies. FIG. 18, bottom panel, is a seriesof graphs comparing the inhibition of various HIV X4 strains againstvarious concentrations of 10E8, 515H7 and 10E8/515H7 antibodies.

FIG. 19, top panel, is a series of graphs comparing inhibition ofvarious HIV strains against varying concentrations of 10E8/Pro140,10E8/iMab, 10E8/515H7 and 10E8/X19 antibodies. FIG. 19, bottom panel,indicates the density of CD4, CCR5 and CXCR4 receptors present on TZM-b1cells.

FIG. 20 compares the binding of CrossMab bispecific antibodies10E8/Pro140, Δ10E8/Pro140 and 4E10/Pro140 to the HIV-1 glycoproteinMPER.

FIG. 21 is a series of graphs comparing the inhibition of variousstrains of HIV against varying concentrations of 4E10, Pro140 and4E10/P140 and 10E8/P140 antibodies.

FIG. 22 is size exclusion chromatography analysis of the CrossMabantibodies 10E8/iMab, 10E8/P140 and 3BNC117/iMab (top panel) and themonoclonal antibodies iMab, 10E8 and Pro140 (bottom panel).

FIG. 23 is size exclusion chromatography analysis of monoclonal antibody10E8 and a chimeric antibody comprised of the 10E8 heavy chain pairedwith the 4E10 light chain.

FIG. 24 is a series of size exclusion chromatography graphs of themonoclonal antibodies 10E8 and 4E10 and a chimeric antibody comprised ofthe 10E8 heavy chain paired with the 4E10 light chain (top panel), themonoclonal antibody 10E8 and 10E8 mutants with potentially stabilizingmutations genetically engineered in the 10E8 light chain (center panel),and the monoclonal antibody 10E8 and 10E8 mutants genetically graftedwith the kappa light chain of non-10E8 antibodies (bottom panel).

FIG. 25 is a size exclusion chromatography graph of the monoclonalantibody 4E10 and 4E10 mutants genetically grafted with the lightregions of 10E8 that included the CDR1 region, CDR2 region, CDR3 region,or combined CDR1, CDR2 and CDR3 regions.

FIG. 26, top panel, is a size exclusion chromatography graph of 10E8chimeric antibodies. CDR123 is a chimeric antibody of the 10E8 heavychain paired with a 10E8 light chain genetically grafted with the 10E8antibody germline CDR region sequences. FW123 is a chimeric antibody ofthe 10E8 heavy chain paired with a 10E8 light chain genetically graftedwith the 10E8 antibody germline framework region sequences. FIG. 26,bottom panel, is a table indicating the expression, HIV MPER bindingability, size exclusion chromatography profile, and HIV neutralizationprofile of the CDR123 and FW123 antibodies.

FIG. 27 is a size exclusion chromatography graph of monoclonal antibody10E8, its somatic variant H6L10, and a CrossMab bispecific antibodyconsisting of H6L10 paired with Pro140.

FIG. 28 is a graph depicting the pharmacokinetics profiles of 10E8,H6L10/Pro 140 and its parental antibodies in a mouse model.

FIG. 29 is a graph comparing the potency of 10E8_(v1.0)/iMab or P140CrossMab antibodies with 10E8/iMab or P140 antibodies.

FIG. 30 is a graph depicting the pharmacokinetics of 10E8 and CrossMabantibodies derived from several 10E8 variants and iMab or P140 in amouse model.

FIGS. 31 is a series of graphs depicting the HIV viral coverage of10E8_(v1.1)/P140 and 10E8_(v2.0)/iMab antibodies (top panel) and sizeexclusion chromatography stability graphs of 10E8_(v1.1)/P140 and10E8_(v2.0)/iMab antibodies(bottom panel).

FIG. 32 is a series of graphs depicting the size exclusion stabilitygraphs of 10E8_(v1.1)/P140 and 10E8_(v2.0)/iMab antibodies stored in PBSat 4° C.

FIG. 33 depicts a native mass spectroscopy analysis of the10E8_(v2.0)/iMab (N297A) antibody.

FIG. 34 is a series of graphs comparing the activity of 10E8_(v1.1)/P140and 10E8_(v2.0)/iMab on a HIV Clade C panel, and the IC50 and IC80activities of the antibodies.

FIGS. 35 and 36 are graphs comparing the potency of 10E8_(v1.1)/P140,10E8_(v2.0)/iMab, and various monoclonal antibodies against HIV.

DETAILED DESCRIPTION

Embodiments of the present invention provide for inhibition of HIV. Invarious implementations, bispecific antibodies are formed, eachincluding heavy chain and light chain components from two differentparent antibodies. One parent antibody specifically binds HIV, forexample, the HIV envelope protein Env. The other parent antibodyspecifically binds a cell membrane protein, for example CD4 and CCR5. Ina bispecific antibody, a heavy chain and light chain from each of twoparental antibodies are combined, providing an antibody in which theantigen binding sites of fragment antigen-binding 1 (Fab1) and Fab2 havedifferent binding specificities. In certain embodiments, the bispecificantibody is a CrossMab format antibody, as shown in FIG. 1. In aCrossMab format, one heavy chain includes a “knob” structure and theother heavy chain includes a corresponding “hole” structure, and thepositions of the constant domains (i.e., CL and CH1) from one parentalantibody are switched, which together ensure correct pairing of heavychains and light chains during assembly.

Various mAbs have been shown to block HIV infection by targeting andbinding to the HIV envelope protein Env (FIGS. 2B and 10). These mAbsinclude, for example, PGT145, PG9, PGT128, PGT121, 10-1074, 3BNC117,VRC01, PGT151, 4E10, and 10E8. In addition, monoclonal antibodies Pro140 (“P140”), Ibalizumab (“iMab”) and 515H7 have been shown to block HIVinfection by targeting and binding to CCR5 , CD4 and CXCR4 human cellmembrane proteins, respectively (FIG. 2A). Specifically, FIG. 2A showshow iMab targets CD4, the primary receptor for HIV-1 entry that isexpressed on human T-cells; and how Pro 140 targets CCR5, a co-receptorfor HIV-1 entry by CCR5 tropic HIV-1. FIG. 2B (adapted fromhttp://www.scripps.edu/news/press/2014/20140424hiv.html) illustrates howthe mAb PGT145 targets the V1/V2 epitope on the HIV viral envelopegp120; how mAb PGT128 targets the glycan on the V3 stem region of HIVgp120; how mAb 3BNC117 targets the CD4 binding site of HIV gp120; howmAb 10E8 targets the membrane proximal external region (MPER) of HIVgp41; and how mAb PGT151 targets an epitope on both HIV gp120 and HIVgp41. In various embodiments, the bispecific antibody (e.g., a HIVCrossMab antibody) of the present invention has the natural architectureof an IgG molecule, but with bispecificity.

Although the ensuing discussion focuses on the use of bispecificantibodies directed to Env and the cell membrane proteins CD4 and CCR5,it is to be understood that this is solely for ease of presentation, andthat any suitable antibody directed to any HIV epitope and any suitableantibody directed to any suitable cell membrane protein may be used andare within the scope of the invention.

Accordingly, in various embodiments, the present invention providesbispecific antibodies that target and bind to the HIV Env protein aswell as the cell membrane proteins CCR5, CD4 and/or CXCR4. In certainembodiments, the bispecific antibodies include sequences (for example,heavy and light chain sequences) derived from, but not limited to, thePGT145, PG9, PGT128, PGT121, 10-1074, 3BNC117, VRC01, PGT151, 4E10,and/or 10E8 antibodies and variants thereof.

The amino acid sequences defining the heavy and light chains of thePGT145 antibody can be found, for example, athttp://www.ncbi.nlm.nih.gov/protein/3U1S_H andhttp://www.ncbi.nlm.nih.gov/protein/3U1S_L, respectively, the entirecontents of which are incorporated herein by reference.

The amino acid sequences defining the heavy and light chains of the PG9antibody can be found, for example, athttp://www.ncbi.nlm.nih.gov/protein/3U4E_H andhttp://www.ncbi.nlm.nih.gov/protein/3MUH_L, respectively, the entirecontents of which are incorporated herein by reference.

The amino acid sequences defining the heavy and light chains of thePGT128 antibody can be found, for example, athttp://www.ncbi.nlm.nih.gov/protein/3TYG_H andhttp://www.ncbi.nlm.nih.gov/protein/3TYG_L, respectively, the entirecontents of which are incorporated herein by reference.

The amino acid sequences defining the heavy and light chains of thePGT121 antibody can be found, for example, athttp://www.ncbi.nlm.nih.gov/protein/4FQC_H andhttp://www.ncbi.nlm.nih.gov/protein/4FQC_L, respectively, the entirecontents of which are incorporated herein by reference.

The amino acid sequences defining the heavy and light chains of the10-1074 antibody can be found, for example, in Mouquet H., et al.,(2012) PNAS, 109(47):E3268-77 (including supplementary information), theentire contents of which are incorporated herein by reference.

The amino acid sequences defining the heavy and light chains of the3BNC117 antibody can be found, for example, athttp://www.ncbi.nlm.nih.gov/protein/4LSV_H andhttp://www.ncbi.nlm.nih.gov/protein/4LSV_L, respectively, the entirecontents of which are incorporated herein by reference.

The amino acid sequences defining the heavy and light chains of theVRC01 antibody can be found, for example, athttp://www.ncbi.nlm.nih.gov/protein/4LST_H andhttp://www.ncbi.nlm.nih.gov/protein/4LST_L, respectively, the entirecontents of which are incorporated herein by reference.

The amino acid sequences defining the heavy and light chains of thePGT151 antibody can be found, for example, athttp://www.ncbi.nlm.nih.gov/protein/4NUG_H andhttp://www.ncbi.nlm.nih.gov/protein/4NUG_L, respectively, the entirecontents of which are incorporated herein by reference.

The amino acid sequences defining the heavy and light chains of the 4E10antibody can be found, for example, athttp://www.ncbi.nlm.nih.gov/protein/4LLV_H andhttp://www.ncbi.nlm.nih.gov/protein/4LLV_L, respectively, the entirecontents of which are incorporated herein by reference.

The amino acid sequences defining the heavy and light chains of the 10E8antibody can be found, for example, athttp://www.ncbi.nlm.nih.gov/protein/4G6F_B andhttp://www.ncbi.nlm.nih.gov/protein/4G6F_D, respectively, the entirecontents of which are incorporated herein by reference.

In certain embodiments, the bispecific antibodies include sequences (forexample, heavy and light chain sequences) derived from, but not limitedto, the P140, iMab (or the MV1 variant) and/or 515H7 antibodies andvariants thereof. The heavy and light chain sequences of the Pro 140,Ibalizumab (or its MV1 variant), and 515H7 antibodies are furtherdescribed, for example, in Olson, W. C. et al., (1999) J Virol.,73(5):4145-55, Trkola, A. et al., (2001) J Virol., 75(2):579-88, U.S.Pat. No. 7,122,185, Burkly L. C. et al., (1992) J Immunol.,149(5):1779-87, Moore J. P. et al., (1992) J Virol., 66(8):4784-93,Reimann K. A., et al., (1997) AIDS Res Hum Retroviruses, 13(11):933-43,International Patent Publication No. WO2014100139, and European PatentPublication No. EP2246364, the entire contents of all of which areincorporated herein by reference.

As used herein, an antibody “variant” refers to an antibody which has anamino acid sequence which differs from the amino acid sequence of aparent antibody from which it is derived. In various embodiments, thevariant has one or more amino acid alterations with respect to theparent antibody.

An embodiment of a bispecific antibody includes a heavy and light chainsequence from the PGT145, PG9, PGT128, PGT121, 10-1074, 3BNC117, VRC01,PGT151, 4E10, or 10E8 antibody or a variant thereof and a heavy andlight chain sequence from the P140, iMab (or the MV1 variant), or 515H7antibody or a variant thereof.

In exemplary embodiments, a series of HIV CrossMab antibodies have beenconstructed including but not limited to, for example, 145/MV1, 117/MV1,128/MV1, 10E8/MV1, 145/P140, 128/P140, 117/P140, 10E8/P140,10E8/alpha-Her2, 10E8/X19, and 4E10/P140. PGT145 (“145”), 3BNC117(“117”), PGT128 (“128”), and 10E8 are four different HIV envelopeantibodies. Pro 140 (“P140”) is a mAb that binds to the cell surfacereceptor CCR5. MV1 is a CD4 antibody that is a modified variant of themAb Ibalizumab (see, for example, International Patent Publication No.WO2014100139, incorporated herein by reference in its entirety). X19 isone of the antibody variants of the anti-cell surface receptor CXCR4(see, for example, U.S. Pat. No. 8,329,178, incorporated herein byreference in its entirety) that does not bind to cells expressing CXCR4(and is therefore used as a non-surface binding control). Alpha-Her2 isa mAb that binds to the Her2 receptor expressed on cells. Many of theseCrossMab antibodies increase the breadth of HIV neutralization ascompared to their parental antibodies (i.e., monoclonal antibodies MV1,145, 117 or 10E8). In addition, many of these antibodies alsosignificantly increase the potency of neutralization against HIV ascompared to their parental antibodies.

The amino acid sequences defining the heavy and light chains of variousHIV CrossMab antibodies are shown below.

145/MV1 antibody:

Amino acid sequence defining the MV1 derived light chain of the 145/MV1antibody —MV1-VLCH1 (SEQ ID NO:1):

DIVMTQSPDSLAVSLGERVTMNCKSSQSLLYSTNQKNYLAWYQQKPGQSPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSVQAEDVAVYYCQQYYSYRTFGGGTKLEIKSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN HKPSNTKVDKKVEPKSC

Amino acid sequence defining the MV1 derived heavy chain of the 145/MV1antibody—MV1-HC-Hole-Cross (SEQ ID NO:2):

QVQLQQSGPEVVKPGASVKMSCKASGYTFTSYVIHWVRQKPGQGLDWIGYINPYNDGTDYDEKFKGKATLTSDTSTSTAYMELSSLRSEDTAVYYCAREKDNYATGAWFAYWGQGTLVTVSSASTAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSL SLSPGK

Amino acid sequence defining the PGT145 derived light chain of the145/MV1 antibody—PGT145-LC (SEQ ID NO:3):

EVVITQSPLFLPVTPGEAASLSCKCSHSLQHSTGANYLAWYLQRPGQTPRLLIHLATHRASGVPDRFSGSGSGTDFTLKISRVESDDVGTYYCMQGLHSPWTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC

Amino acid sequence defining the PGT145 derived heavy chain of the145/MV1 antibody—PGT145-HC-Knob (SEQ ID NO:4):

QVQLVQSGAEVKKPGSSVKVSCKASGNSFSNHDVHWVRQATGQGLEWMGWMSHEGDKTGLAQKFQGRVTITRDSGASTVYMELRGLTADDTAIYYCLTGSKHRLRDYFLYNEYGPNYEEWGDYLATLDVWGHGTAVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV LHEALHSHYTQKSLSLSPGK

117/MV1 antibody:

Amino acid sequence defining the MV1 derived light chain of the 117/MV1antibody —MV1-VLCH1 (SEQ ID NO:1):

DIVMTQSPDSLAVSLGERVTMNCKSSQSLLYSTNQKNYLAWYQQKPGQSPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSVQAEDVAVYYCQQYYSYRTFGGGTKLEIKSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN HKPSNTKVDKKVEPKSC

Amino acid sequence defining the MV1 derived heavy chain of the 117/MV1antibody—MV1-HC-Hole-Cross (SEQ ID NO:2):

QVQLQQSGPEVVKPGASVKMSCKASGYTFTSYVIHWVRQKPGQGLDWIGYINPYNDGTDYDEKFKGKATLTSDTSTSTAYMELSSLRSEDTAVYYCAREKDNYATGAWFAYWGQGTLVTVSSASTAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSL SLSPGK

Amino acid sequence defining the 3BNC117 derived light chain of the117/MV1 antibody—3BNC117-LC (SEQ ID NO:5):

DIQMTQSPSSLSASVGDTVTITCQANGYLNWYQQRRGKAPKLLIYDGSKLERGVPSRFSGRRWGQEYNLTINNLQPEDIATYFCQVYEFVVFGQGTKVQVDIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGEC

Amino acid sequence defining the 3BNC117 derived heavy chain of the117/MV1 antibody—3BNC117-HC-Knob (SEQ ID NO:6):

QVQLLQSGAAVTKPGASVRVSCEASGYNIRDYFIHWWRQAPGQGLQWVGWINPKTGQPNNPRQFQGRVSLTRHASWDFDTFSFYMDLKALRSDDTAVYFCARQRSDYWDFDVWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLS PGK

128/MV1 antibody:

Amino acid sequence defining the MV1 derived light chain of the 128/MV1antibody —MV1-VLCH1 (SEQ ID NO:1):

DIVMTQSPDSLAVSLGERVTMNCKSSQSLLYSTNQKNYLAWYQQKPGQSPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSVQAEDVAVYYCQQYYSYRTFGGGTKLEIKSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN HKPSNTKVDKKVEPKSC

Amino acid sequence defining the MV1 derived heavy chain of the 128/MV1antibody—MV1-HC-Hole-Cross (SEQ ID NO:2):

QVQLQQSGPEVVKPGASVKMSCKASGYTFTSYVIHWVRQKPGQGLDWIGYINPYNDGTDYDEKFKGKATLTSDTSTSTAYMELSSLRSEDTAVYYCAREKDNYATGAWFAYWGQGTLVTVSSASTAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSL SLSPGK

Amino acid sequence defining the PGT128 derived light chain of the128/MV1 antibody—PGT128-LC (SEQ ID NO:7):

QSALTQPPSASGSPGQSITISCTGTSNNFVSWYQQHAGKAPKLVIYDVNKRPSGVPDRFSGSKSGNTASLTVSGLQTDDEAVYYCGSLVGNWDVIFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGST VEKTVAPTECS

Amino acid sequence defining the PGT128 derived heavy chain of the128/MV1 antibody—PGT128-HC-Knob (SEQ ID NO:8):

QPQLQESGPTLVEASETLSLTCAVSGDSTAACNSFWGWVRQPPGKGLEWVGSLSHCASYWNRGWTYHNPSLKSRLTLALDTPKNLVFLKLNSVTAADTATYYCARFGGEVLRYTDWPKPAWVDLWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEAL HSHYTQKSLSLSPGK

10E8/MV1 antibody:

Amino acid sequence defining the MV1 derived light chain of the 10E8/MV1antibody—MV1-VLCH1 (SEQ ID NO:1):

DIVMTQSPDSLAVSLGERVTMNCKSSQSLLYSTNQKNYLAWYQQKPGQSPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSVQAEDVAVYYCQQYYSYRTFGGGTKLEIKSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN HKPSNTKVDKKVEPKSC

Amino acid sequence defining the MV1 derived heavy chain of the 10E8/MV1antibody—MV1-HC-Hole-Cross (SEQ ID NO:2):

QVQLQQSGPEVVKPGASVKMSCKASGYTFTSYVIHWVRQKPGQGLDWIGYINPYNDGTDYDEKFKGKATLTSDTSTSTAYMELSSLRSEDTAVYYCAREKDNYATGAWFAYWGQGTLVTVSSASTAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSL SLSPGK

Amino acid sequence defining the 10E8 derived light chain of the10E8/MV1 antibody—10E8-LC (SEO ID NO:91:

YELTQETGVSVALGRTVTITCRGDSLRSHYASWYQKKPGQAPILLFYGKNNRPSGVPDRFSGSASGNRASLTISGAQAEDDAEYYCSSRDKSGSRLSVFGGGTKLTVLSQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHE GSTVXKTVAPTECS

Amino acid sequence defining the 10E8 derived heavy chain of the10E8/MV1 antibody—10E8-HC-Knob (SEQ ID NO:10):

EVQLVESGGGLVKPGGSLRLSCSASGFDFDNAWMTWVRQPPGKGLEWVGRITGPGEGWSVDYAAPVEGRFTISRLNSINFLYLEMNNLRMEDSGLYFCARTGKYYDFWSGYPPGEEYFQDWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHY TQKSLSLSPGK

Δ10E8/MV1 antibody

Amino acid sequence defining the MV1 derived light chain of theΔ10E8/MV1 antibody MV1-VLCH1 (SEQ ID NO:1):

DIVMTQSPDSLAVSLGERVTMNCKSSQSLLYSTNQKNYLAWYQQKPGQSPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSVQAEDVAVYYCQQYYSYRTFGGGTKLEIKSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN HKPSNTKVDKKVEPKSC

Amino acid sequence defining the MV1 derived heavy chain of theΔ10E8/MV1 antibody MV1-HC-Hole-Cross (SEQ ID NO:2):

QVQLQQSGPEVVKPGASVKMSCKASGYTFTSYVIHWVRQKPGQGLDWIGYINPYNDGTDYDEKFKGKATLTSDTSTSTAYMELSSLRSEDTAVYYCAREKDNYATGAWFAYWGQGTLVTVSSASTAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSL SLSPGK

Amino acid sequence defining the Δ 10E8 derived light chain of theΔ10E8/MV1 antibody Δ10E8-LC (SEQ ID NO:21):

YELTQETGVSVALGRTVTITCRGDSLRSHYASWYQKKPGQAPILLFYGKNNRPSGVPDRFSGASGNRASLTISGAQAEDDAEYYCSSRDKSGSRLSVFGGGTKLTVLSQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEG STVEKTVAPTECS

Amino acid sequence defining the Δ 10E8 derived heavy chain of theΔ10E8/MV1 antibody 10E8-HC-Knob (SEQ ID NO:22):

EVQLVESGGGLVKPGGSLRLSCSASGFDFDNAWMTWVRQPPGKGLEWVGRITGPGEGWSVDYAAPVEGRFTISRLNSINFLYLEMNNLRMEDSGLYFCARTGKYYDFWSGYPPGEEYFQDWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHY TQKSLSLSPGK

151/MV1 antibody

Amino acid sequence defining the MV1 derived light chain of the 151/MV1antibody —MV1-VLCH1 (SEQ ID NO:1):

DIVMTQSPDSLAVSLGERVTMNCKSSQSLLYSTNQKNYLAWYQQKPGQSPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSVQAEDVAVYYCQQYYSYRTFGGGTKLEIKSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN HKPSNTKVDKKVEPKSC

Amino acid sequence defining the MV1 derived heavy chain of the 151/MV1antibody MV1-HC-Hole-Cross (SEQ ID NO:2):

QVQLQQSGPEVVKPGASVKMSCKASGYTFTSYVIHWVRQKPGQGLDWIGYINPYNDGTDYDEKFKGKATLTSDTSTSTAYMELSSLRSEDTAVYYCAREKDNYATGAWFAYWGQGTLVTVSSASTAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSL SLSPGK

Amino acid sequence defining the PGT151 derived light chain of the151/MV1 antibody PGT151-LC (SEQ ID NO:23):

DIVMTQTPLSLSVTPGQPASISCKSSESLRQSNGKTSLYWYRQKPGQSPQLLVFEVSNRFSGVSDRFVGSGSGTDFTLRISRVEAEDVGFYYCMQSKDFPLTFGGGTKVDLKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC

Amino acid sequence defining the PGT151 derived heavy chain of the151/MV1 antibody PGT151-HC-Knob (SEQ ID NO:24):

RVQLVESGGGVVQPGKSVRLSCVVSDFPFSKYPMYWVRQAPGKGLEWVAAISGDAWHVVYSNSVQGRFLVSRDNVKNTLYLEMNSLKIEDTAVYRCARMFQESGPPRLDRWSGRNYYYYSGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEAL HSHYTQKSLSLSPGK

145/P140 antibody:

Amino acid sequence defining the Pro 140 derived light chain of the145/P140 antibody—PRO140-VLCH1 (SEQ ID NO:11):

DIVMTQSPLSLPVTPGEPASISCRSSQRLLSSYGHTYLHWYLQKPGQSPQLLIYEVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCSQSTHVPLTFGQGTKVEIKSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN HKPSNTKVDKKVEPKSC

Amino acid sequence defining the Pro 140 derived heavy chain of the145/P140 antibody—PRO140-HC-Hole-Cross (SEQ ID NO:12):

EVQLVESGGGLVKPGGSLRLSCAASGYTFSNYWIGWVRQAPGKGLEWIGDIYPGGNYIRNNEKFKDKTTLSADTSKNTAYLQMNSLKTEDTAVYYCGSSFGSNYVFAWFTYWGQGTLVTVSSASTAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSL SLSPGK

Amino acid sequence defining the PGT145 derived light chain of the145/P140 antibody—PGT145-LC (SEQ ID NO:3):

EVVITQSPLFLPVTPGEAASLSCKCSHSLQHSTGANYLAWYLQRPGQTPRLLIHLATHRASGVPDRFSGSGSGTDFTLKISRVESDDVGTYYCMQGLHSPWTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC

Amino acid sequence defining the PGT145 derived heavy chain of the145/P140 antibody—PGT145-HC-Knob (SEQ ID NO:4):

QVQLVQSGAEVKKPGSSVKVSCKASGNSFSNHDVHWVRQATGQGLEWMGWMSHEGDKTGLAQKFQGRVTITRDSGASTVYMELRGLTADDTAIYYCLTGSKHRLRDYFLYNEYGPNYEEWGDYLATLDVWGHGTAVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV LHEALHSHYTQKSLSLSPGK

117/P140 antibody:

Amino acid sequence defining the Pro 140 derived light chain of the117/P140 antibody—PRO140-VLCH1 (SEQ ID NO:11):

DIVMTQSPLSLPVTPGEPASISCRSSQRLLSSYGHTYLHWYLQKPGQSPQLLIYEVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCSQSTHVPLTFGQGTKVEIKSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN HKPSNTKVDKKVEPKSC

Amino acid sequence defining the Pro 140 derived heavy chain of the117/P140 antibody—PRO140-HC-Hole-Cross (SEQ ID NO:12):

EVQLVESGGGLVKPGGSLRLSCAASGYTFSNYWIGWVRQAPGKGLEWIGDIYPGGNYIRNNEKFKDKTTLSADTSKNTAYLQMNSLKTEDTAVYYCGSSFGSNYVFAWFTYWGQGTLVTVSSASTAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSL SLSPGK

Amino acid sequence defining the 3BNC117 derived light chain of the117/P140 antibody—3BNC117-LC (SEQ ID NO:5):

DIQMTQSPSSLSASVGDTVTITCQANGYLNWYQQRRGKAPKLLIYDGSKLERGVPSRFSGRRWGQEYNLTINNLQPEDIATYFCQVYEFVVFGQGTKVQVDIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGEC

Amino acid sequence defining the 3BNC117 derived heavy chain of the117/P140 antibody—3BNC117-HC-Knob (SEQ ID NO:6):

QVQLLQSGAAVTKPGASVRVSCEASGYNIRDYFIHWWRQAPGQGLQWVGWINPKTGQPNNPRQFQGRVSLTRHASWDFDTFSFYMDLKALRSDDTAVYFCARQRSDYWDFDVWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLS PGK

128/P140 antibody:

Amino acid sequence defining the Pro 140 derived light chain of the128/P140 antibody—PRO140-VLCH1 (SEQ ID NO:11):

DIVMTQSPLSLPVTPGEPASISCRSSQRLLSSYGHTYLHWYLQKPGQSPQLLIYEVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCSQSTHVPLTFGQGTKVEIKSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN HKPSNTKVDKKVEPKSC

Amino acid sequence defining the Pro 140 derived heavy chain of the128/P140 antibody—PRO140-HC-Hole-Cross (SEQ ID NO:12):

EVQLVESGGGLVKPGGSLRLSCAASGYTFSNYWIGWVRQAPGKGLEWIGDIYPGGNYIRNNEKFKDKTTLSADTSKNTAYLQMNSLKTEDTAVYYCGSSFGSNYVFAWFTYWGQGTLVTVSSASTAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSL SLSPGK

Amino acid sequence defining the PGT128 derived light chain of the128/P140 antibody—PGT128-LC (SEQ ID NO:7):

QSALTQPPSASGSPGQSITISCTGTSNNFVSWYQQHAGKAPKLVIYDVNKRPSGVPDRFSGSKSGNTASLTVSGLQTDDEAVYYCGSLVGNWDVIFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGST VEKTVAPTECS

Amino acid sequence defining the PGT128 derived heavy chain of the128/P140 antibody—PGT128-HC-Knob (SEQ ID NO:8):

QPQLQESGPTLVEASETLSLTCAVSGDSTAACNSFWGWVRQPPGKGLEWVGSLSHCASYWNRGWTYHNPSLKSRLTLALDTPKNLVFLKLNSVTAADTATYYCARFGGEVLRYTDWPKPAWVDLWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEAL HSHYTQKSLSLSPGK

10E8/P140 antibody:

Amino acid sequence defining the Pro 140 derived light chain of the10E8/P140 antibody—PRO140-VLCH1 (SEQ ID NO:11):

DIVMTQSPLSLPVTPGEPASISCRSSQRLLSSYGHTYLHWYLQKPGQSPQLLIYEVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCSQSTHVPLTFGQGTKVEIKSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN HKPSNTKVDKKVEPKSC

Amino acid sequence defining the Pro 140 derived heavy chain of the10E8/P140 antibody—PRO140-HC-Hole-Cross (SEQ ID NO:12):

EVQLVESGGGLVKPGGSLRLSCAASGYTFSNYWIGWVRQAPGKGLEWIGDIYPGGNYIRNNEKFKDKTTLSADTSKNTAYLQMNSLKTEDTAVYYCGSSFGSNYVFAWFTYWGQGTLVTVSSASTAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSL SLSPGK

Amino acid sequence defining the 10E8 derived light chain of the10E8/P140 antibody—10E8-LC (SEQ ID NO:9):

YELTQETGVSVALGRTVTITCRGDSLRSHYASWYQKKPGQAPILLFYGKNNRPSGVPDRFSGSASGNRASLTISGAQAEDDAEYYCSSRDKSGSRLSVFGGGTKLTVLSQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHE GSTVXKTVAPTECS

Amino acid sequence defining the 10E8 derived heavy chain of the10E8/P140 antibody—10E8-HC-Knob (SEQ ID NO:10):

EVQLVESGGGLVKPGGSLRLSCSASGFDFDNAWMTWVRQPPGKGLEWVGRITGPGEGWSVDYAAPVEGRFTISRLNSINFLYLEMNNLRMEDSGLYFCARTGKYYDFWSGYPPGEEYFQDWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHY TQKSLSLSPGK

Δ10E8/P140 antibody [000119] Amino acid sequence defining the PRO140derived light chain of the Δ10E8/P140 antibody—PRO140-VLCH1 (SEQ IDNO:11):

DIVMTQSPLSLPVTPGEPASISCRSSQRLLSSYGHTYLHWYLQKPGQSPQLLIYEVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCSQSTHVPLTFGQGTKVEIKSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN HKPSNTKVDKKVEPKSC

Amino acid sequence defining the PRO140 derived heavy chain of theΔ10E8/P140 antibody—PRO140-Hole-Cross (SEQ ID NO:12):

EVQLVESGGGLVKPGGSLRLSCAASGYTFSNYWIGWVRQAPGKGLEWIGDIYPGGNYIRNNEKFKDKTTLSADTSKNTAYLQMNSLKTEDTAVYYCGSSFGSNYVFAWFTYWGQGTLVTVSSASTAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSL SLSPGK

Amino acid sequence defining the 410E8 derived light chain of theΔ10E8/P140 antibody—410E8-LC (SEQ ID NO:21):

YELTQETGVSVALGRTVTITCRGDSLRSHYASWYQKKPGQAPILLFYGKNNRPSGVPDRFSGASGNRASLTISGAQAEDDAEYYCSSRDKSGSRLSVFGGGTKLTVLSQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEG STVXKTVAPTECS

Amino acid sequence defining the 410E8 derived heavy chain of theΔ10E8/P140 antibody—10E8-HC-Knob (SEQ ID NO:22):

EVQLVESGGGLVKPGGSLRLSCSASGFDFDNAWMTWVRQPPGKGLEWVGRITGPGEGWSVDYAAPVEGRFTISRLNSINFLYLEMNNLRMEDSGLYFCARTGKYYDFWSGYPPGEEYFQDWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHY TQKSLSLSPGK

151/P140 antibody

Amino acid sequence defining the PRO140 derived light chain of the151/P140 antibody—PRO140-VLCH1 (SEQ ID NO:11):

DIVMTQSPLSLPVTPGEPASISCRSSQRLLSSYGHTYLHWYLQKPGQSPQLLIYEVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCSQSTHVPLTFGQGTKVEIKSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN HKPSNTKVDKKVEPKSC

Amino acid sequence defining the PRO140 derived heavy chain of the151/P140 antibody—PRO140-Hole-Cross (SEQ ID NO:12):

EVQLVESGGGLVKPGGSLRLSCAASGYTFSNYWIGWVRQAPGKGLEWIGDIYPGGNYIRNNEKFKDKTTLSADTSKNTAYLQMNSLKTEDTAVYYCGSSFGSNYVFAWFTYWGQGTLVTVSSASTAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSL SLSPGK

Amino acid sequence defining the PGT151 derived light chain of the151/P140 antibody—PGT151-LC (SEQ ID NO:23):

DIVMTQTPLSLSVTPGQPASISCKSSESLRQSNGKTSLYWYRQKPGQSPQLLVFEVSNRFSGVSDRFVGSGSGTDFTLRISRVEAEDVGFYYCMQSKDFPLTFGGGTKVDLKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC

Amino acid sequence defining the PGT151 derived heavy chain of the151/P140 antibody—PGT151-HC-Knob (SEQ ID NO:24):

RVQLVESGGGVVQPGKSVRLSCVVSDFPFSKYPMYWVRQAPGKGLEWVAAISGDAWHVVYSNSVQGRFLVSRDNVKNTLYLEMNSLKIEDTAVYRCARMFQESGPPRLDRWSGRNYYYYSGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEAL HSHYTQKSLSLSPGK

10E8/Alpha-Her2 antibody:

Amino acid sequence defining the alpha-Her2 derived light chain of the10E8/Alpha-Her2 antibody—antiHer2-VLCH1 (SEQ ID NO:13):

DIVMTQSHKFMSTSVGDRVSITCKASQDVNTAVAWYQQKPGHSPKLLIYSASFRYTGVPDRFTGNRSGTDFTFTISSVQAEDLAVYYCQQHYTTPPTFGGGTKVEIKSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSN TKVDKKVEPKSC

Amino acid sequence defining the alpha-Her2 derived heavy chain of the10E8/Alpha-Her2 antibody—antiHer2-HC-Hole-Cross (SEQ ID NO:14):

QVQLQQSGPELVKPGASLKLSCTASGFNIKDTYIHWVKQRPEQGLEWIGRIYPTNGYTRYDPKFQDKATITADTSSNTAYLQVSRLTSEDTAVYYCSRWGGDGFYAMDYWGQGASVTVSSASTAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSL SPGK

Amino acid sequence defining the 10E8 derived light chain of the10E8/Alpha-Her2 antibody—10E8-LC (SEQ ID NO:9):

YELTQETGVSVALGRTVTITCRGDSLRSHYASWYQKKPGQAPILLFYGKNNRPSGVPDRFSGSASGNRASLTISGAQAEDDAEYYCSSRDKSGSRLSVFGGGTKLTVLSQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHE GSTVXKTVAPTECS

Amino acid sequence defining the 10E8 derived heavy chain of the10E8/Alpha-Her2 antibody—10E8-HC-Knob (SEQ ID NO:10):

EVQLVESGGGLVKPGGSLRLSCSASGFDFDNAWMTWVRQPPGKGLEWVGRITGPGEGWSVDYAAPVEGRFTISRLNSINFLYLEMNNLRMEDSGLYFCARTGKYYDFWSGYPPGEEYFQDWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHY TQKSLSLSPGK

4E10/P140 antibody:

Amino acid sequence defining the Pro 140 derived light chain of the4E10/P140 antibody—PRO140-VLCH1 (SEQ ID NO:11):

DIVMTQSPLSLPVTPGEPASISCRSSQRLLSSYGHTYLHWYLQKPGQSPQLLIYEVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCSQSTHVPLTFGQGTKVEIKSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN HKPSNTKVDKKVEPKSC

Amino acid sequence defining the Pro 140 derived heavy chain of the4E10/P140 antibody—PRO140-HC-Hole-Cross (SEQ ID NO:12):

EVQLVESGGGLVKPGGSLRLSCAASGYTFSNYWIGWVRQAPGKGLEWIGDIYPGGNYIRNNEKFKDKTTLSADTSKNTAYLQMNSLKTEDTAVYYCGSSFGSNYVFAWFTYWGQGTLVTVSSASTAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSL SLSPGK

Amino acid sequence defining the 4E10 derived light chain of the4E10/P140 antibody—4E10-LC (SEQ ID NO:17):

EIVLTQSPGTQSLSPGERATLSCRASQSVGNNKLAWYQQRPGQAPRLLIYGASSRPSGVADRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGQSLSTFGQGTKVEVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC

Amino acid sequence defining the 4E10 derived heavy chain of the4E10/P140 antibody—PGT145-HC-Knob (SEQ ID NO:18):

QVQLVQSGAEVKRPGSSVTVSCKASGGSFSTYALSWVRQAPGRGLEWMGGVIPLLTITNYAPRFQGRITITADRSTSTAYLELNSLRPEDTAVYYCAREGTTGAGWLGKPIGAFAHWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKS LSLSPGK

10E8/X19 antibody:

Amino acid sequence defining the X19 derived light chain of the 10E8/X19antibody —X19-VLCH1 (SEQ ID NO:19):

EIVLTQSPATLSVSPGRRATLSCRASQSVNTNLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQHYGSSPLTFGGGTKLEIKSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSN TKVDKKVEPKSC

Amino acid sequence defining the X19 derived heavy chain of the 10E8/X19antibody—X19-HC-Hole-Cross (SEQ ID NO:20):

QVQLVQSGGGVVQPGRSLRLSCAASGFTFSSYPMHWVRQAPGKGLEWMTVISSDGRNKYYPDSVKGRFTISRDNSKNTLYLQMNSLRPEDTAVYYCARGGYHDFWSGPDYWGQGTLVTVSSASTAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLS LSPGK

Amino acid sequence defining the 10E8 derived light chain of the10E8/X19 antibody —10E8-LC (SEQ ID NO:9):

YELTQETGVSVALGRTVTITCRGDSLRSHYASWYQKKPGQAPILLFYGKNNRPSGVPDRFSGSASGNRASLTISGAQAEDDAEYYCSSRDKSGSRLSVFGGGTKLTVLSQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHE GSTVXKTVAPTECS

Amino acid sequence defining the 10E8 derived heavy chain of the10E8/X19 antibody—PGT145-HC-Knob (SEQ ID NO:10):

EVQLVESGGGLVKPGGSLRLSCSASGFDFDNAWMTWVRQPPGKGLEWVGRITGPGEGWSVDYAAPVEGRFTISRLNSINFLYLEMNNLRMEDSGLYFCARTGKYYDFWSGYPPGEEYFQDWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHY TQKSLSLSPGK

10E8/515H7 antibody

Amino acid sequence defining the 515H7 derived light chain of the10E8/515H7 antibody—515H7-VLCH1 (SEQ ID NO:25):

DIVMSQSPSSLAVSAGEKVTMSCKSSQSLFNSRTRKNYLAWYQQKPGQSPKLLIYWASARDSGVPARFTGSGSETYFTLTISRVQAEDLAVYYCMQSFNLRTFGGGTKLEIKASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHK PSNTKVDKKVEPKSC

Amino acid sequence defining the 515H7 derived heavy chain of the10E8/515H7 antibody—515H7-Hole-Cross (SEQ ID NO:26):

EVNLVESGGGLVQPGGSLRLSCATSGFTFTDNYMSWVRQPPGKALEWLGFIRNKANGYTTDYSASVRGRFTISRDNSQSILYLQMNALRAEDSATYYCARDVGSNYFDYWGQGTTLTVSSASTAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSL SPGK

Amino acid sequence defining the 10E8 derived light chain of the10E8/515H7 antibody—10E8-LC (SEO ID NO:91:

YELTQETGVSVALGRTVTITCRGDSLRSHYASWYQKKPGQAPILLFYGKNNRPSGVPDRFSGSASGNRASLTISGAQAEDDAEYYCSSRDKSGSRLSVFGGGTKLTVLSQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHE GSTVEKTVAPTECS

Amino acid sequence defining the 10E8 derived heavy chain of the10E8/515H7 antibody—10E8-HC-Knob (SEQ ID NO:10):

EVQLVESGGGLVKPGGSLRLSCSASGFDFDNAWMTWVRQPPGKGLEWVGRITGPGEGWSVDYAAPVEGRFTISRLNSINFLYLEMNNLRMEDSGLYFCARTGKYYDFWSGYPPGEEYFQDWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHY TQKSLSLSPGK

Chimeric CDR123 antibody (SEQ ID NO:27):

SELTQDPAVSVALGQTVRITCRGDSLRSHYASWYQQKPGQAPVLVIYGKNNRPSGIPDRFSGSSSGNTASLTITGAQAEDEADYYCSSRDKSGSRLSVFGGGTKLTVLSQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHE GSTVEKTVAPTECS

Chimeric FW123 (SEQ ID NO:28):

YELTQETGVSVALGRTVTITCQGDSLRSYYASWYQKKPGQAPILLFYGKNNRPSGVPDRFSGSASGNRASLTISGAQAEDDAEYYCNSRDSSGNHLVVFGGGTKLTVLSQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHE GSTVEKTVAPTECS

10E8V1.0/iMab antibody

Amino acid sequence defining the MV1 derived light chain of the10E8v1.0/MV1 antibody MV1-VLCH1 (SEQ ID NO:1):

DIVMTQSPDSLAVSLGERVTMNCKSSQSLLYSTNQKNYLAWYQQKPGQSPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSVQAEDVAVYYCQQYYSYRTFGGGTKLEIKSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN HKPSNTKVDKKVEPKSC

Amino acid sequence defining the MV1 derived heavy chain of the10E8v1.0/MV1 antibody MV1-HC-Hole-Cross (SEQ ID NO:2):

QVQLQQSGPEVVKPGASVKMSCKASGYTFTSYVIHWVRQKPGQGLDWIGYINPYNDGTDYDEKFKGKATLTSDTSTSTAYMELSSLRSEDTAVYYCAREKDNYATGAWFAYWGQGTLVTVSSASTAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSL SLSPGK

Amino acid sequence defining the 10E8v1.0 derived light chain of the10E8v1.0/iMab antibody—10E8v1.0-LC (SEQ ID NO:29):

ASELTQDPAVSVALKQTVTITCRGDSLRSHYVSWYQKKPGQAPVLVFYGKNNRPSGIPDRFSGSSSGNTASLTIAGAQAEDDADYYCSSRDKSGSRLSVFGGGTKLTVLSQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTH EGSTVEKTVAPTECS

Amino acid sequence defining the 10E8v1.0 derived heavy chain of the10E8v1.0/iMab antibody—10E8v1.0-HC-Knob (SEQ ID NO:30):

EVRLVESGGGLVKPGGSLRLSCSASGFNFDDAWMTWVRQPPGKGLEWVGRISGPGEGWSVDYAESVKGRFTISRLNSINFLYLEMNNVRTEDTGYYFCARTGKHYDFWSGYPPGEEYFQDWGQGTLVIVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHY TQKSLSLSPGK

10E8V1.1/iMab antibody

Amino acid sequence defining the MV1 derived light chain of the10E8v1.1/iMab antibody MV1-VLCH1 (SEQ ID NO:1):

DIVMTQSPDSLAVSLGERVTMNCKSSQSLLYSTNQKNYLAWYQQKPGQSPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSVQAEDVAVYYCQQYYSYRTFGGGTKLEIKSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN HKPSNTKVDKKVEPKSC

Amino acid sequence defining the MV1 derived heavy chain of the10E8v1.1/iMab antibody MV1-HC-Hole-Cross (SEQ ID NO:2):

QVQLQQSGPEVVKPGASVKMSCKASGYTFTSYVIHWVRQKPGQGLDWIGYINPYNDGTDYDEKFKGKATLTSDTSTSTAYMELSSLRSEDTAVYYCAREKDNYATGAWFAYWGQGTLVTVSSASTAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSL SLSPGK

Amino acid sequence defining the 10E8v1.1 derived light chain of the10E8v1.1/iMab antibody—10E8v1.1-LC (SEQ ID NO:31):

ASELTQDPAVSVALKQTVTITCRGDSLRSHYVSWYQKKPGQAPVLVFYGKNNRPSGIPDRFSGSSSGNTASLTIAGAQAEDDADYYCSSRDKSGSRLSVFGGGTKLTVLSQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTH EGSTVEKTVAPTECS

Amino acid sequence defining the 10E8v1.1 derived heavy chain of the10E8v1.1/iMab antibody—10E8v1.1 HC-Knob (SEQ ID NO:32):

EVRLVESGGGLVKPGGSLRLSCSASGFNFDDAWMTWVRQPPGKGLEWVGRISGPGEGWSVDYAESVKGRFTISRLNSINFLYLEMNNVRTEDTGYYFCARTGKYYDFWSGYPPGEEYFQDWGQGTLVIVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHY TQKSLSLSPGK

10E8V2.0/iMab antibody

Amino acid sequence defining the MV1 derived light chain of the10E8v2.0/iMab antibody MV1-VLCH1 (SEQ ID NO:1):

DIVMTQSPDSLAVSLGERVTMNCKSSQSLLYSTNQKNYLAWYQQKPGQSPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSVQAEDVAVYYCQQYYSYRTFGGGTKLEIKSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN HKPSNTKVDKKVEPKSC

Amino acid sequence defining the MV1 derived heavy chain of the10E8v2.0/iMab antibody MV1-HC-Hole-Cross (SEQ ID NO:2):

QVQLQQSGPEVVKPGASVKMSCKASGYTFTSYVIHWVRQKPGQGLDWIGYINPYNDGTDYDEKFKGKATLTSDTSTSTAYMELSSLRSEDTAVYYCAREKDNYATGAWFAYWGQGTLVTVSSASTAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSL SLSPGK

Amino acid sequence defining the 10E8v2.0 derived light chain of the10E8v2.0/iMab antibody—10E8v2.0-LC (SEQ ID NO:33):

ASELTQDPAVSVALKQTVTITCRGDSLRSHYASWYQKKPGQAPILLFYGKNNRPSGVPDRFSGSASGNRASLTISGAQAEDDAEYYCSSRDKSGSRLSVFGGGTKLTVLSQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTH EGSTVEKTVAPTECS

Amino acid sequence defining the 10E8v2.0 derived heavy chain of the10E8v2.0/iMab antibody—10E8v2.0-HC-Knob (SEQ ID NO:34):

EVRLVESGGGLVKPGGSLRLSCSASGFNFDDAWMTWVRQPPGKGLEWVGRISGPGEGWSVDYAESVKGRFTISRLNSINFLYLEMNNVRTEDTGYYFCARTGKHYDFWSGYPPGEEYFQDWGQGTLVIVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHY TQKSLSLSPGK

10E8V3.0/iMab antibody

Amino acid sequence defining the MV1 derived light chain of the10E8v3.0/iMab antibody MV1-VLCH1 (SEQ ID NO:1):

DIVMTQSPDSLAVSLGERVTMNCKSSQSLLYSTNQKNYLAWYQQKPGQSPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSVQAEDVAVYYCQQYYSYRTFGGGTKLEIKSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN HKPSNTKVDKKVEPKSC

Amino acid sequence defining the MV1 derived heavy chain of the10E8v3.0/iMab antibody MV1-HC-Hole-Cross (SEQ ID NO:2):

QVQLQQSGPEVVKPGASVKMSCKASGYTFTSYVIHWVRQKPGQGLDWIGYINPYNDGTDYDEKFKGKATLTSDTSTSTAYMELSSLRSEDTAVYYCAREKDNYATGAWFAYWGQGTLVTVSSASTAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSL SLSPGK

Amino acid sequence defining the 10E8v3.0 derived light chain of the10E8v3.0/iMab antibody—10E8v3.0-LC (SEQ ID NO:35):

SELTQETGVSVALGRTVTITCRGDSLRSHYASWYQKKPGQAPILLFYGKNNRPSGIHDRFSGSASGNRASLTISGAQAEDDAEYYCSSRDKSGSRLSVFGGGTKLTVLSQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHE GSTVEKTVAPTECS

Amino acid sequence defining the 10E8v3.0 derived heavy chain of the10E8v3.0/iMab antibody—10E8v3.0-HC-Knob (SEQ ID NO:36):

EVQLVESGGDLVKPGGSLRLSCSASGFSFKNTWMTWVRQAPGKGLEWVGRITGPGEGWTSDYAATVQGRFTISRNNMIDMLYLEMNRLRTDDTGLYYCVHTEKYYNFWGGYPPGEEYFQHWGRGTLVIVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHY TQKSLSLSPGK

10E8V1.0/P140 (H6L10/PRO140) antibody

[000171] Amino acid sequence defining the PRO140 derived light chain ofthe 10E8V1.0/P140 antibody—PRO140-VLCH1 (SEQ ID NO:11):

DIVMTQSPLSLPVTPGEPASISCRSSQRLLSSYGHTYLHWYLQKPGQSPQLLIYEVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCSQSTHVPLTFGQGTKVEIKSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN HKPSNTKVDKKVEPKSC

Amino acid sequence defining the PRO140 derived heavy chain of the10E8V1.0/P140 antibody—PRO140-Hole-Cross (SEQ ID NO:12):

EVQLVESGGGLVKPGGSLRLSCAASGYTFSNYWIGWVRQAPGKGLEWIGDIYPGGNYIRNNEKFKDKTTLSADTSKNTAYLQMNSLKTEDTAVYYCGSSFGSNYVFAWFTYWGQGTLVTVSSASTAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSL SLSPGK

Amino acid sequence defining the L10 derived light chain of the10E8V1.0/P140 antibody—L10-LC (SEQ ID NO:29):

ASELTQDPAVSVALKQTVTITCRGDSLRSHYVSWYQKKPGQAPVLVFYGKNNRPSGIPDRFSGSSSGNTASLTIAGAQAEDDADYYCSSRDKSGSRLSVFGGGTKLTVLSQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTH EGSTVEKTVAPTECS

Amino acid sequence defining the H6 derived heavy chain of the10E8V1.0/P140 antibody—H6-HC-Knob (SEQ ID NO:30):

EVRLVESGGGLVKPGGSLRLSCSASGFNFDDAWMTWVRQPPGKGLEWVGRISGPGEGWSVDYAESVKGRFTISRLNSINFLYLEMNNVRTEDTGYYFCARTGKHYDFWSGYPPGEEYFQDWGQGTLVIVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHY TQKSLSLSPGK

10E8V1.1/P140 antibody [000176] Amino acid sequence defining the PRO140derived light chain of the 10E8v1.1/P140 antibody PRO140-VLCH1 (SEQ IDNO:11):

DIVMTQSPLSLPVTPGEPASISCRSSQRLLSSYGHTYLHWYLQKPGQSPQLLIYEVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCSQSTHVPLTFGQGTKVEIKSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN HKPSNTKVDKKVEPKSC

Amino acid sequence defining the P140 derived heavy chain of the10E8v1.1/P140 antibody PRO 140-HC-Hole-Cross (SEQ ID NO:12):

EVQLVESGGGLVKPGGSLRLSCAASGYTFSNYWIGWVRQAPGKGLEWIGDIYPGGNYIRNNEKFKDKTTLSADTSKNTAYLQMNSLKTEDTAVYYCGSSFGSNYVFAWFTYWGQGTLVTVSSASTAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSL SLSPGK

Amino acid sequence defining the 10E8v1.1 derived light chain of the10E8v1.1/P140 antibody—10E8v1.1-LC (SEQ ID NO:31):

ASELTQDPAVSVALKQTVTITCRGDSLRSHYVSWYQKKPGQAPVLVFYGKNNRPSGIPDRFSGSSSGNTASLTIAGAQAEDDADYYCSSRDKSGSRLSVFGGGTKLTVLSQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTH EGSTVEKTVAPTECS

Amino acid sequence defining the 10E8v1.1 derived heavy chain of the10E8v1.1/P140 antibody—10E8v1.1 HC-Knob (SEQ ID NO:32):

EVRLVESGGGLVKPGGSLRLSCSASGFNFDDAWMTWVRQPPGKGLEWVGRISGPGEGWSVDYAESVKGRFTISRLNSINFLYLEMNNVRTEDTGYYFCARTGKYYDFWSGYPPGEEYFQDWGQGTLVIVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHY TQKSLSLSPGK

10E8V2.0/P140 antibody

Amino acid sequence defining the PRO140 derived light chain of the10E8v2.0/P140 antibody PRO140-VLCH1 (SEQ ID NO:11):

DIVMTQSPLSLPVTPGEPASISCRSSQRLLSSYGHTYLHWYLQKPGQSPQLLIYEVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCSQSTHVPLTFGQGTKVEIKSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN HKPSNTKVDKKVEPKSC

Amino acid sequence defining the P140 derived heavy chain of the10E8v2.0/P140 antibody PRO140-HC-Hole-Cross (SEQ ID NO:12):

EVQLVESGGGLVKPGGSLRLSCAASGYTFSNYWIGWVRQAPGKGLEWIGDIYPGGNYIRNNEKFKDKTTLSADTSKNTAYLQMNSLKTEDTAVYYCGSSFGSNYVFAWFTYWGQGTLVTVSSASTAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSL SLSPGK

Amino acid sequence defining the 10E8v2.0 derived light chain of the10E8v2.0/P140 antibody—10E8v2.0-LC (SEQ ID NO:33):

ASELTQDPAVSVALKQTVTITCRGDSLRSHYASWYQKKPGQAPILLFYGKNNRPSGVPDRFSGSASGNRASLTISGAQAEDDAEYYCSSRDKSGSRLSVFGGGTKLTVLSQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTH EGSTVEKTVAPTECS

Amino acid sequence defining the 10E8v2.0 derived heavy chain of the10E8v2.0/P140 antibody—10E8v2.0 HC-Knob (SEQ ID NO:34):

EVRLVESGGGLVKPGGSLRLSCSASGFNFDDAWMTWVRQPPGKGLEWVGRISGPGEGWSVDYAESVKGRFTISRLNSINFLYLEMNNVRTEDTGYYFCARTGKHYDFWSGYPPGEEYFQDWGQGTLVIVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHY TQKSLSLSPGK

10E8V3.0/P140 antibody

Amino acid sequence defining the PRO140 derived light chain of the10E8v3.0/P140 antibody PRO140-VLCH1 (SEQ ID NO:11):

DIVMTQSPLSLPVTPGEPASISCRSSQRLLSSYGHTYLHWYLQKPGQSPQLLIYEVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCSQSTHVPLTFGQGTKVEIKSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN HKPSNTKVDKKVEPKSC

Amino acid sequence defining the P140 derived heavy chain of the10E8v3.0/P140 antibody PRO140-HC-Hole-Cross (SEQ ID NO:12):

EVQLVESGGGLVKPGGSLRLSCAASGYTFSNYWIGWVRQAPGKGLEWIGDIYPGGNYIRNNEKFKDKTTLSADTSKNTAYLQMNSLKTEDTAVYYCGSSFGSNYVFAWFTYWGQGTLVTVSSASTAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSL SLSPGK

Amino acid sequence defining the 10E8v3.0 derived light chain of the10E8v3.0/P140 antibody—10E8v3.0-LC (SEQ ID NO:35):

SELTQETGVSVALGRTVTITCRGDSLRSHYASWYQKKPGQAPILLFYGKNNRPSGIHDRFSGSASGNRASLTISGAQAEDDAEYYCSSRDKSGSRLSVFGGGTKLTVLSQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHE GSTVEKTVAPTECS

Amino acid sequence defining the 10E8v3.0 derived heavy chain of the10E8v3.0/P140 antibody—10E8v3.0 HC-Knob (SEQ ID NO:36):

EVQLVESGGDLVKPGGSLRLSCSASGFSFKNTWMTWVRQAPGKGLEWVGRITGPGEGWTSDYAATVQGRFTISRNNMIDMLYLEMNRLRTDDTGLYYCVHTEKYYNFWGGYPPGEEYFQHWGRGTLVIVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHY TQKSLSLSPGK

In various embodiments, at least one of the heavy chain and/or lightchain sequences derived from the PGT145, PG9, PGT128, PGT121, 10-1074,3BNC117, VRC01, PGT151, 4E10, 10E8, P140, iMab (or the MV1 variant),515H7 antibodies and variants thereof are paired together to form abispecific antibody (e.g., a HIV CrossMab antibody). In an exemplaryembodiment, at least one of the disclosed heavy and light chainsselected from SEQ ID NOs: 1-36 are paired together to form a bispecificantibody (e.g., a HIV CrossMab antibody).

In various embodiments, the amino acid sequence of the bispecificantibody (e.g., HIV CrossMab antibody) further includes an amino acidanalog, an amino acid derivative, or other non-classical amino acids.

In various embodiments, the bispecific antibody (e.g., HIV CrossMabantibody) comprises a sequence that is at least 60% identical to awild-type heavy or light chain sequence of the PGT145, PG9, PGT128,PGT121, 10-1074, 3BNC117, VRC01, PGT151, 4E10, or 10E8 antibody. Invarious embodiments, the bispecific antibody (e.g., HIV CrossMabantibody) comprises a sequence that is at least 60% identical to awild-type heavy chain or light chain sequence of the P140, iMab (or theMV1 variant), or 515H7 antibody. In exemplary embodiments, thebispecific antibody (e.g., HIV CrossMab antibody) comprises a sequencethat is at least 60% identical to SEQ ID NOs: 1-36.

In various embodiments, the bispecific antibody (e.g., HIV CrossMabantibody) may comprise a sequence that is at least about 60%, at leastabout 61%, at least about 62%, at least about 63%, at least about 64%,at least about 65%, at least about 66%, at least about 67%, at leastabout 68%, at least about 69%, at least about 70%, at least about 71%,at least about 72%, at least about 73%, at least about 74%, at leastabout 75%, at least about 76%, at least about 77%, at least about 78%,at least about 79%, at least about 80%, at least about 81%, at leastabout 82%, at least about 83%, at least about 84%, at least about 85%,at least about 86%, at least about 87%, at least about 88%, at leastabout 89%, at least about 90%, at least about 91%, at least about 92%,at least about 93%, at least about 94%, at least about 95%, at leastabout 96%, at least about 97%, at least about 98%, at least about 99%,or 100% identical to a wild-type heavy chain or light chain sequence ofthe PGT145, PG9, PGT128, PGT121, 10-1074, 3BNC117, VRC01, PGT151, 4E10,or 10E8 antibody.

In various embodiments, the bispecific antibody (e.g., HIV CrossMabantibody) may comprise a sequence that is at least about 60%, at leastabout 61%, at least about 62%, at least about 63%, at least about 64%,at least about 65%, at least about 66%, at least about 67%, at leastabout 68%, at least about 69%, at least about 70%, at least about 71%,at least about 72%, at least about 73%, at least about 74%, at leastabout 75%, at least about 76%, at least about 77%, at least about 78%,at least about 79%, at least about 80%, at least about 81%, at leastabout 82%, at least about 83%, at least about 84%, at least about 85%,at least about 86%, at least about 87%, at least about 88%, at leastabout 89%, at least about 90%, at least about 91%, at least about 92%,at least about 93%, at least about 94%, at least about 95%, at leastabout 96%, at least about 97%, at least about 98%, at least about 99%,or 100% identical to a wild-type heavy chain or light chain sequence ofthe P140, iMab (or the MV1 variant), or 515H7 antibody.

In exemplary embodiments, the bispecific antibody (e.g., HIV CrossMabantibody) may comprise a sequence that is at least about 60%, at leastabout 61%, at least about 62%, at least about 63%, at least about 64%,at least about 65%, at least about 66%, at least about 67%, at leastabout 68%, at least about 69%, at least about 70%, at least about 71%,at least about 72%, at least about 73%, at least about 74%, at leastabout 75%, at least about 76%, at least about 77%, at least about 78%,at least about 79%, at least about 80%, at least about 81%, at leastabout 82%, at least about 83%, at least about 84%, at least about 85%,at least about 86%, at least about 87%, at least about 88%, at leastabout 89%, at least about 90%, at least about 91%, at least about 92%,at least about 93%, at least about 94%, at least about 95%, at leastabout 96%, at least about 97%, at least about 98%, at least about 99%,or 100% identical to SEQ ID NOs: 1-36.

Homology or identity may be determined in various ways that are withinthe skill in the art, for instance, using publicly available computersoftware such as BLAST, BLAST-2, ALIGN or Megalign (DNASTAR) software.BLAST (Basic Local Alignment Search Tool) analysis using the algorithmemployed by the programs blastp, blastn, blastx, tblastn and tblastx(Karlin et al., (1990) PROC. NATL. ACAD. SCI. USA 87, 2264-2268;Altschul, (1993) J. MOL. EVOL. 36, 290-300; Altschul et al., (1997)NUCLEIC ACIDS RES. 25, 3389-3402, incorporated by reference) aretailored for sequence similarity searching. The approach used by theBLAST program is to first consider similar segments between a querysequence and a database sequence, then to evaluate the statisticalsignificance of all matches that are identified and finally to summarizeonly those matches which satisfy a preselected threshold ofsignificance. For a discussion of basic issues in similarity searchingof sequence databases see Altschul et al., (1994) NATURE GENETICS 6,119-129 which is fully incorporated by reference. Those skilled in theart can determine appropriate parameters for measuring alignment,including any algorithms needed to achieve maximal alignment over thefull length of the sequences being compared. The search parameters forhistogram, descriptions, alignments, expect (i.e., the statisticalsignificance threshold for reporting matches against databasesequences), cutoff, matrix and filter are at the default settings. Thedefault scoring matrix used by blastp, blastx, tblastn, and tblastx isthe BLOSUM62 matrix (Henikoff et al., (1992) PROC. NATL. ACAD. SCI. USA89, 10915-10919, fully incorporated by reference). Four blastnparameters may be adjusted as follows: Q=10 (gap creation penalty); R=10(gap extension penalty); wink=1 (generates word hits at everywink.sup.th position along the query); and gapw=16 (sets the windowwidth within which gapped alignments are generated). The equivalentBlastp parameter settings may be Q=9; R=2; wink=1; and gapw=32. Searchesmay also be conducted using the NCBI (National Center for BiotechnologyInformation) BLAST Advanced Option parameter (e.g.: —G, Cost to open gap[Integer]: default =5 for nucleotides/11 for proteins; —E, Cost toextend gap [Integer]: default =2 for nucleotides/ 1 for proteins; —q,Penalty for nucleotide mismatch [Integer]: default =−3; -r, reward fornucleotide match [Integer]: default =1; —e, expect value [Real]: default=10; -W, wordsize [Integer]: default =11 for nucleotides/ 28 formegablast/ 3 for proteins; —y, Dropoff (X) for blast extensions in bits:default =20 for blastn/ 7 for others; −X, X dropoff value for gappedalignment (in bits): default =15 for all programs, not applicable toblastn; and −Z, final X dropoff value for gapped alignment (in bits): 50for blastn, 25 for others). ClustalW for pairwise protein alignments mayalso be used (default parameters may include, e.g., Blosum62 matrix andGap Opening Penalty =10 and Gap Extension Penalty =0.1). A Bestfitcomparison between sequences, available in the GCG package version 10.0,uses DNA parameters GAP=50 (gap creation penalty) and LEN=3 (gapextension penalty) and the equivalent settings in protein comparisonsare GAP=8 and LEN=2.

In various embodiments, the bispecific antibody (e.g., HIV CrossMabantibody) comprises a sequence that includes at least one amino acidalteration with respect to a wild-type heavy or light chain sequence ofthe PGT145, PG9, PGT128, PGT121, 10-1074, 3BNC117, VRC01, PGT151, 4E10,or 10E8 antibody. In various embodiments, the bispecific antibody (e.g.,HIV CrossMab antibody) comprises a sequence that includes at least oneamino acid alteration with respect to a wild-type heavy or light chainsequence of the P140, iMab (or the MV1 variant), 515H7 antibody. Inexemplary embodiments, the bispecific antibody (e.g., HIV CrossMabantibody) comprises a sequence that includes at least one amino acidalteration with respect to SEQ ID NOs: 1-36.

In various embodiments, the bispecific antibody (e.g., HIV CrossMabantibody) comprises a sequence that includes at least about 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23,24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41,42, 43, 44, 45, 46, 47, 40, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59,60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77,78, 79, or 80 amino acid alterations with respect to a wild-type heavyor light chain sequence of the PGT145, PG9, PGT128, PGT121, 10-1074,3BNC117, VRC01, PGT151, 4E10, or 10E8 antibody.

In various embodiments, the bispecific antibody (e.g., HIV CrossMabantibody) comprises a sequence that includes at least about 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23,24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41,42, 43, 44, 45, 46, 47, 40, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59,60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77,78, 79, or 80 amino acid alterations with respect to a wild-type heavyor light chain sequence of the P140, iMab (or the MV1 variant), or 515H7antibody.

In exemplary embodiments, the bispecific antibody (e.g., HIV CrossMabantibody) comprises a sequence that includes at least about 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23,24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41,42, 43, 44, 45, 46, 47, 40, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59,60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77,78, 79, or 80 amino acid alterations with respect to SEQ ID NOs: 1-36.

The amino acid alteration can be an amino acid deletion, insertion,substitution, or modification. In one embodiment, the amino acidalteration is an amino acid deletion. In another embodiment, the aminoacid alteration is an amino acid substitution.

In various embodiments, the amino acid alteration may be in theComplementarity Determining Regions (CDRs) of the bispecific antibody(e.g., the CDR1, CDR2 or CDR3 regions). In another embodiment, the aminoacid alteration may be in the framework regions (FWs) of the bispecificantibody (e.g., the FW1, FW2, FW3, or FW4 regions). In a furtherembodiment, the amino acid alteration may be in the joining regions (Jregions) of the bispecific antibody (e.g., the J1, J2, J3, J4, J5, J6,or J7 regions).

Also provided herein are chimeric antibody derivatives of the bispecificantibodies, i.e., antibody molecules in which a portion of the heavyand/or light chain is identical with or homologous to correspondingsequences in antibodies derived from a particular species or belongingto a particular antibody class or subclass, while the remainder of thechain(s) is identical with or homologous to corresponding sequences inantibodies derived from another species or belonging to another antibodyclass or subclass, as well as fragments of such antibodies, so long asthey exhibit the desired biological activity. For example, thebispecific antibody may include a heavy and/or light chain in which oneor more CDRs or FWs derived from an antibody selected from a PGT145,PG9, PGT128, PGT121, 10-1074, 3BNC117, VRC01, PGT151, 4E10, 10E8, P140,iMab (or the MV1 variant), or 515H7 antibody are replaced with one ormore CDRs or FWs derived from a different antibody selected from aPGT145, PG9, PGT128, PGT121, 10-1074, 3BNC117, VRC01, PGT151, 4E10,10E8, P140, iMab (or the MV1 variant), or 515H7 antibody.

Modification of the amino acid sequence of recombinant binding proteinis achieved using any known technique in the art e.g., site-directedmutagenesis or PCR based mutagenesis. Such techniques are described, forexample, in Sambrook et al., Molecular Cloning: A Laboratory Manual,Cold Spring Harbor Press, Plainview, N.Y., 1989 and Ausubel et al.,Current Protocols in Molecular Biology, John Wiley & Sons, New York,N.Y., 1989.

Methods for producing antibodies, such as those disclosed herein, areknown in the art. For example, DNA molecules encoding light chainvariable regions and/or heavy chain variable regions can be chemicallysynthesized using the sequence information provided herein. SyntheticDNA molecules can be ligated to other appropriate nucleotide sequences,including, e.g., expression control sequences, to produce conventionalgene expression constructs encoding the desired antibodies. Productionof defined gene constructs is within routine skill in the art.Alternatively, the sequences provided herein can be cloned out ofhybridomas by conventional hybridization techniques or polymerase chainreaction (PCR) techniques, using synthetic nucleic acid probes whosesequences are based on sequence information provided herein, or priorart sequence information regarding genes encoding the heavy and lightchains.

Nucleic acids encoding desired antibodies can be incorporated (ligated)into expression vectors, which can be introduced into host cells throughconventional transfection or transformation techniques. Exemplary hostcells are E.coli cells, Chinese hamster ovary (CHO) cells, humanembryonic kidney 293 (HEK 293) cells, HeLa cells, baby hamster kidney(BHK) cells, monkey kidney cells (COS), human hepatocellular carcinomacells (e.g., Hep G2), and myeloma cells that do not otherwise produceIgG protein. Transformed host cells can be grown under conditions thatpermit the host cells to express the genes that encode theimmunoglobulin light and/or heavy chain variable regions. Specificexpression and purification conditions will vary depending upon theexpression system employed.

In various embodiments, the bispecific antibodies of the presentinvention (e.g., HIV CrossMab antibodies) are used in therapy. Forexample, the bispecific antibody (e.g., HIV CrossMab antibody) can beused to neutralize HIV in a mammal (e.g., a human patient). For example,antibodies of the invention can bind to HIV so as to partially orcompletely inhibit one or more biological activities of the virus. In anembodiment, the bispecific antibody (e.g., HIV CrossMab antibody)neutralizes a R5-tropic HIV. In another embodiment, the bispecificantibody (e.g., HIV CrossMab antibody) neutralizes a X4-tropic HIV. In afurther embodiment, the bispecific antibody (e.g., HIV CrossMabantibody) neutralizes a R5X4 dual-tropic HIV. In some embodiments, useof the antibody to neutralize HIV in a mammal comprises administering tothe mammal a therapeutically effective amount of the antibody.

Generally, a therapeutically effective amount of active component is inthe range of, for example, about 0.1 mg/kg to about 100 mg/kg, e.g.,about 1 mg/kg to about 100 mg/kg, e.g., about 1 m/kg to about 10 mg/kgof the body weight of the patient. In various embodiments, atherapeutically effective amount of active component is in a range ofabout 0.01 mg/kg to about 10 mg/kg of the body weight of the patient,for example, about 0.01 mg/kg, about 0.02 mg/kg, about 0.03 mg/kg, about0.04 mg/kg, about 0.05 mg/kg, about 0.06 mg/kg, about 0.07 mg/kg, about0.08 mg/kg, about 0.09 mg/kg, about 0.1 mg/kg, about 0.2 mg/kg, about0.3 mg/kg, about 0.4 mg/kg, about 0.5 mg/kg, about 0.6 mg/kg, about 0.7mg/kg, about 0.8 mg/kg, about 0.9 mg/kg, about 1 mg/kg, about 1.1 mg/kg,about 1.2 mg/kg, about 1.3 mg/kg, about 1.4 mg/kg, about 1.5mg/kg, about1.6 mg/kg, about 1.7 mg/kg, about 1.8 mg/kg, 1.9 mg/kg, about 2 mg/kg,about 3 mg/kg, about 4 mg/kg, about 5 mg/kg, about 6 mg/kg, about 7mg/kg, about 8 mg/kg, about 9 mg/kg, about 10 mg/kg body weight,inclusive of all values and ranges therebetween.

The amount administered will depend on variables such as the type andextent of disease or indication to be treated, the overall health of thepatient, the in vivo potency of the antibody, the pharmaceuticalformulation, and the route of administration. The initial dosage can beincreased beyond the upper level in order to rapidly achieve the desiredblood-level or tissue level. Alternatively, the initial dosage can besmaller than the optimum, and the dosage may be progressively increasedduring the course of treatment. Human dosage can be optimized, e.g., ina conventional Phase I dose escalation study designed to run from, forexample, 0.5 mg/kg to 20 mg/kg. Dosing frequency can vary, depending onfactors such as route of administration, dosage amount and the diseasebeing treated. Exemplary dosing frequencies are more than once daily,about once per day, about twice a day, about three times a day, aboutfour times a day, about five times a day, about every other day, aboutevery third day, about once a week, about once every two weeks, aboutonce every month, about once every two months, about once every threemonths, about once every six months, or about once every year.Formulation of antibody-based drugs is within ordinary skill in the art.

For therapeutic use, an antibody may be combined with a pharmaceuticallyacceptable carrier. As used herein, “pharmaceutically acceptablecarrier” means buffers, carriers, and excipients suitable for use incontact with the tissues of human beings and animals without excessivetoxicity, irritation, allergic response, or other problem orcomplication, commensurate with a reasonable benefit/risk ratio. Thecarrier(s) should be “acceptable” in the sense of being compatible withthe other ingredients of the formulations and not deleterious to therecipient. Pharmaceutically acceptable carriers include buffers,solvents, dispersion media, coatings, isotonic and absorption delayingagents, and the like, that are compatible with pharmaceuticaladministration. The use of such media and agents for pharmaceuticallyactive substances is known in the art.

Pharmaceutical compositions containing antibodies, such as thosedisclosed herein, can be presented in a dosage unit form and can beprepared by any suitable method. A pharmaceutical composition should beformulated to be compatible with its intended route of administration.Examples of routes of administration are intravenous (IV), intradermal,inhalation, transdermal, topical, transmucosal, and rectaladministration. In an embodiment, the route of administration forantibodies of the invention is IV infusion.

Useful formulations can be prepared by methods well known in thepharmaceutical art. For example, see Remington's PharmaceuticalSciences, 18th ed. (Mack Publishing Company, 1990). Formulationcomponents suitable for parenteral administration include a sterilediluent such as water for injection, saline solution, fixed oils,polyethylene glycols, glycerine, propylene glycol or other syntheticsolvents; antibacterial agents such as benzyl alcohol or methyl paraben;antioxidants such as ascorbic acid or sodium bisulfite; chelating agentssuch as EDTA; buffers such as acetates, citrates or phosphates; andagents for the adjustment of tonicity such as sodium chloride ordextrose. For intravenous administration, suitable carriers includephysiological saline, bacteriostatic water, Cremophor ELTM (BASF,Parsippany, N.J.) or phosphate buffered saline (PBS). The carrier shouldbe stable under the conditions of manufacture and storage, and should bepreserved against microorganisms. The carrier can be a solvent ordispersion medium containing, for example, water, ethanol, polyol (forexample, glycerol, propylene glycol, and liquid polyetheylene glycol),and suitable mixtures thereof.

Pharmaceutical formulations preferably are sterile. Sterilization can beaccomplished, for example, by filtration through sterile filtrationmembranes. Where the composition is lyophilized, filter sterilizationcan be conducted prior to or following lyophilization andreconstitution.

FIGS. 13 and 14 demonstrate that some iMab-based CrossMabs have greaterpotency and breadth than parental Abs. Except otherwise stated, alliMab-based bispecific antibodies were constructed using the MV1 variant.IC80, the antibody concentration that confers 80% neutralization ofviral infectivity, is one method to evaluate antibody potency againstHIV. The lower the IC80 number (indicated in the y-axis of the graphs interm of antibody concentration (μg/ml)), the more potent the antibody isat neutralizing a particular HIV strain or isolate. IC50, the antibodyconcentration that confers 50% neutralization of viral infectivity, isanother method to evaluate antibody potency against HIV. The lower theIC50 number (indicated in the y-axis of the graphs in term of antibodyconcentration (μg/ml)), the more potent the antibody is at neutralizinga particular HIV strain or isolate.

Various sets of antibodies were tested against a large panel of HIV-1pseudoviruses (118 different HIV viral isolates) representative of HIVenvelope diversity by geography, clade, tropism, and stage of infection.IC80 and IC50 were used to evaluate the strength of antiviral potencyand breadth. The bottom right panels in FIGS. 13 and 14 clearlydemonstrate that, as compared to the parental antibodies iMab and 10E8,the bispecific CrossMab of the two together (10E8/iMab) neutralizesalmost all HIV viruses (each virus is indicated as a dot) more potently.The other antibody sets (used to make 145/iMab, 117/iMab, 128/iMab and151/iMab) sometimes enhance HIV potency compared to their parentalcomponents and sometimes do not.

As shown in FIG. 15, the antibody iMab is also relatively potent incell-cell neutralizing assays. PGT145, 3BNC117, 10E8, PGT128 and PGT151are relatively potent at neutralizing cell-free viral infection, but arepoor in neutralizing viruses in cell-cell transmission assays. Creatingbispecific antibodies including PGT145, 3BNC117, 10E8, PGT128 and PGT151with iMab makes these chimeric antibodies active at neutralizing virusesin a cell-cell transmission assay. It can be seen that 10E8/iMab is themost potent antibody in these comparative studies. It is also found that10E8/iMab is most active in preventing cell-cell transmission in vitro.

As illustrated in FIG. 16, the improved potency of 10E8/iMab isstatistically significant. FIG. 3 shows that improved potency requirescovalent linkage of the antibody, i.e., the CrossMab format (sinceco-administration of two parental antibodies, iMab and 10E8, provides alower MPI than the fused and physically linked bispecific 10E8/iMabantibody). FIGS. 10-14 provide further evidence of the improved potencyof iMab-derived CrossMab antibodies over its parental antibodies.

In summary, it is found that, for the iMab-based CrossMabs (fused withPGT145, 3BNC117, PGT151, PGT128 and 10E8), 117/iMab improves breadth butnot potency; 145/iMab, 151/iMab and 128/iMab improve breadth andpotency; and 10E8/iMab markedly improves breadth and potency. In termsof epitope location/accessibility and potential models ofneutralization, 10E8/iMab appears to exhibit pre- and post-attachmentneutralization; 145/iMab, 151/iMab and 117/iMab appear to exhibitpre-attachment neutralization; and 117/iMab may show signs of stericrestriction and potentially reduced potency for some viruses. 10E8/iMabalso exhibits potent activity against HIV cell-to-cell transmission.

As also shown in the top panels and bottom left panels of FIGS. 13 and14, Pro 140-based CrossMab activities are sometimes weaker than theirparental antibodies and corresponding iMab-based CrossMabs, as shown bythe high concentrations required to reach IC80 and IC50. Anchoring ofthese four mAbs to the host cell receptor CCR5 via another host cellreceptor-binding antibody called Pro 140 does not improve the antiviralpotency or breadth (as measured by IC80 against a large panel of HIVisolates) compared to their respective parental antibodies. These panelsindicate that Pro140-based CrossMabs for these four antibodies areweaker than their corresponding iMab-based CrossMabs (IC50 and IC80comparisons of Pro140-based vs. iMab-based CrossMabs).

As shown in the bottom right panels of FIGS. 13 and 14, 10E8/P140, afifth Pro 140-based CrossMab, is more potent than its parentalantibodies and 10E8/iMab CrossMab. These panels illustrate a comparisonof the potency (IC80 or IC50) of parental mAb Pro140 (right-most columnof data points in these panels), bispecific CrossMab 10E8/P140 (secondfrom right column of data points in these panels), and parental mAb 10E8(center column of data points in these panels) against a large panel ofHIV isolates. These panels also illustrate a comparison of the potency(IC80 or IC50) of parental mAb iMab (left-most column of data points inthese panels), bispecific CrossMab 10E8/iMab (second from left column ofdata points in these panels), and parental mAb 10E8 (center column ofdata points in these panels) against a large panel of HIV isolates. Thesecond from left and second from right columns of data points in thesepanels illustrate a comparison of the potency (IC80 or IC50) of thebispecific CrossMabs 10E8/iMab and 10E8/P140 against a large panel ofHIV isolates.

Pro 140 is known to not have activity against X4 HIV viruses, as X4viruses use CXCR4 as a co-receptor for HIV-1 entry, and Pro 140 binds toCCR5. 10E8 alone has very weak activity against X4 viruses. However, thebispecific CrossMab 10E8/Pro 140 can neutralize all X4 viruses tested todate better than either of the parent antibodies. The panels of FIG. 4illustrate the effectiveness of 10E8, Pro 140, and 10E8/P140 bispecificCrossMab antibody in inhibition of various strains of HIV.

As shown in FIG. 5, 10E8/Pro140 CrossMab is a more potent inhibitor ofvarious strains of HIV than the co-administration of the two parentalantibodies, demonstrating a synergistic, not merely additive,enhancement of potency with this particular bispecific antibody.

As shown in FIG. 6, a CrossMab of 10E8 fused to a non-membrane boundantibody (X19) does not provide enhanced potency, as can be seen whencompared to membrane bound 10E8/P140. Thus, the potency of the 10E8/P140CrossMab appears to require anchoring of 10E8 to the cell membrane.However, membrane binding alone does not afford the enhanced potency ofthese CrossMabs. FIG. 7 shows that anchoring 10E8 on HER2 does notprovide substantial potency enhancement as compared to anchoring 10E8 onCCR5. Anchoring of 10E8 to a viral receptor specifically (in this caseCCR5 via Pro 140 or CD4 via iMab) provides enhanced antiviral activity.

Δ10E8 is a mutant version of the 10E8 mAb that has a one amino aciddeletion in the light chain FR3. Compared to 10E8, Δ10E8 has a muchweaker epitope binding activity, as illustrated in FIG. 8A and panels Aand B in FIG. 8B. However, once the Δ10E8 was anchored on a cellreceptor (by combining Δ10E8 and iMab in a CrossMab antibody-iMabspecifically binds cell receptor CD4), FIG. 8B, panels C and D, showthat its inhibition activity is improved. These data suggest thecontribution of specific cell receptor anchoring, i.e., anchoring on aviral receptor or a viral co-receptor, in enhancing the activity of thisHIV antibody. Still, while Δ10E8/P140 CrossMab has improved antiviralactivity over Δ10E8, it is still not as potent as 10E8/P140 CrossMab.Δ10E8/P140 CrossMab is comparatively more effective in neutralizing R5viruses than it is in neutralizing X4 viruses.

4E10 is an anti-gp4l MPER mAb known to be less potent than the anti-gp4lMPER mAb 10E8. Similar to the results for Δ10E8, FIGS. 20 and 21 showthat anchoring 4E10 on co-receptor CCR5 (via Pro 140 in a CrossMabantibody) enhanced antiviral activity of 4E10 significantly. Takentogether, this suggests that the anchoring of a number of anti-gp41 MPERAbs to either CCR5 or CD4 (via combining the MPER Abs with P140 or iMabin a CrossMab antibody bispecific) can greatly improve the potency andbreadth of the respective anti-gp41 MPER Ab.

Multiple parameters contribute to enhanced activity of certainbispecific CrossMabs against HIV, including parental Ab potency,affinity, and pre- and post-attachment neutralization abilities. Inparticular, the 10E8/Pro140 CrossMab represents an effective combinationin terms of overcoming energetic, spatial and temporal constraints,targeting sequential/interdependent steps in the entry process, epitopelocation/accessibility, binding affinity, pre-and post-attachmentneutralization, and binding geometry. As shown in FIG. 20, 4E10/Pro140has a greater binding affinity for MPER than Δ10E8/Pro140 and10E8/Pro140. FIG. 9 shows the inhibition potency of 10E8/Pro140,Δ10E8/Pro140 and 4E10/Pro140, and their parental antibodies 10E8, Δ10E8and 4E10 against various strains of HIV. FIGS. 10 and 13-17 provideadditional evidence of the greater potency of CrossMab antibodies ascompared to their parental antibodies individually and the parentalantibodies in combination.

The enhanced antiviral coverage of 10E8/iMab and 10E8/Pro140 CrossMabsis illustrated in FIG. 10, which depict the potency and breadth ofseveral antibodies against HIV. The x-axis indicates the concentrationof a particular antibody, the y-axis indicates the percent of a largepanel of HIV viral isolates neutralized by a particular antibody at aspecific concentration, and each line indicates a different antibodyevaluated. The left-most lines along the x-axis and those that canclosely approach or reach 100% on the y-axis indicate a highly potentand broad antibody against HIV. 10E8/P140 CrossMab and 10E8/iMabCrossMab are among the most effective antibodies with respect to bothviral coverage and potency, and are significantly more effective thantheir parental antibodies.

FIGS. 18 and 19 show the potency of the CrossMab 10E8/515H7 antibody ascompared to its parental antibodies and previously discussed antibodies.The potency of a CrossMab antibody does not appear to correlate directlywith the density of cell membrane protein targets, as the density ofCCR5 (the target of Pro140) is less than that of CD4 (the target ofibalizumab), yet the potency of 10E8/Pro140-derived CrossMab antibody isgreater than that of 10E8/iMab-derived CrossMab antibody.

As shown in FIG. 22, the lack of single, sharp peaks in size exclusionchromatography indicates a type of instability indicative of multiplemolecular species for 10E8 and 10E8-derived CrossMab antibodies. Table 1recites various process and formulation modifications used to resolvethe 10E8 instability. However, as indicated by the “X” in the SEC orSize Exclusion Chromatography column, the modifications wereunsuccessful in providing a single, sharp peak.

TABLE 1 Process and formulation screen to resolve 10E8 instabilityConditions SEC Purpose EDTA* X sequester metal ions, ↓ enzymaticactivity Acetic Acid* X ↓ pH, stabilize protonated form of free thiois,↓ reduction activity L Lysine* X competitive inhibitor against reductioncomponents CuSO4* X maintain reducing components in oxidized form,enzyme inhibitor 3-day X decreased cell death, ↓ enzymatic activityharvesting SEC running X modification of analytical conditions buffercondition His formulation X modification of analytical conditions buffer

Pairing the 10E8 heavy chain with the light chain of 4E10 resolves theinstability issue, as shown in FIG. 23, producing a functional, thoughless potent, antibody. This result indicates that the instability of10E8 is due to the light chain. Various modifications of the 10E8 lightchain, shown in the center and bottom panels of FIG. 24, such as removalof a C-terminal serine, engineering a lambda-variable region andkappa-constant region chimera, engineering an additional disulfide bondbetween the 10E8 heavy and light chains, or genetically grafting kappalight chain regions of non-10E8 antibodies onto the 10E8 light chain donot fully resolve 10E8 instability. As shown in FIGS. 25 and 26, theinstability is likely due to a combination of the ComplementarityDetermining Regions (“CDRs”) and the framework regions (“FWs”) of 10E8.Using 10E8-HC/4E10-LC, each 10E8 light chain CDR was grafted into 4E1OLCindividually or in concert, as shown in FIG. 25. Addition of 10E8 LCCDR2 and CDR3 are well tolerated, but addition of 10E8 LC CDR1 disruptsthe single peak. When all 10E8 CDRs are grafted onto 4E10, the peak isbroad. Grafting 10E8 CDRs or frameworks onto its germline light chain Xresults in a single peak, as shown in FIG. 26, but effectiveness in MPERbinding and HIV neutralization is decreased. Table 2 summarizes the 10E8light chain variants tested and the efficacy thereof.

TABLE 2 Generated 10E8 LC variants Modification Expression MPER bindingSEC Neutralization* λLC′ → ΔS

X

λLC → κLC

X

LC CDR grafting

↓ ? (κLC Ab1) LC CDR grafting

↓ ? (κLC Ab2) H—L S—S bond X X 10E8-H/4E10-L

↓

↓ 10E8-H/4E10-L X X CDR1 (10E8) 10E8-H/4E10-L

↓

↓ CDR2 (10E8) 10E8-H/4E10-L

↓

↓ CDR3 (10E8) 10E8-H/4E10-L

↓ ? ↓ CDR123 (10E8)

Variant H6L10 of 10E8 antibody was found to be active, non-autoreactive,and stable by size exclusion chromatography, as shown in FIG. 27.H6L10/Pro 140 and its parental antibodies were found to have comparablepharmacokinetics profiles in mice, as shown in FIG. 28. However, asshown in FIG. 29, the H6L10 variant of 10E8 (referred to as 10E8_(v1.0))combined with P140 in a bispecific antibody is substantially less potentthan 10E8/P140 when tested against a large panel of HIV strains. TheH6L10 variant of 10E8 (referred to as 10E8_(v1.0)) combined with iMab ina bispecific antibody retains the same relative amount of potency ascompared to 10E8/iMab when tested against a large panel of HIV strains,but 10E8_(v1.0)/iMab possesses the same instability as 10E8/iMab asdetermined by size exclusion chromatography and indicated by an X inTable 3. In an embodiment, the H6L10 variant may further include a S74Wmutation.

Table 3 below lists exemplary variants, their activities, size exclusionchromatography results, and pharmacokinetics (“PK”) results.

As indicated above, 10E8_(V1.0) is a somatic variant of 10E8 known asH6L10. As a mAb, H6L10 has a single peak by SEC but reduced activitycompared to 10E8. H6L10/Pro140 CrossMab has single SEC peak and goodmouse PK, but reduced anti-HIV activity. H6L10/iMab CrossMab has doubleSEC peaks and poor mouse PK, but its activity against HIV is roughly thesame as 10E8/iMab. 10E8_(V1.1) includes a single point mutation inH6L10. When paired with Pro140 in a CrossMab bispecific, this constructhas single SEC peak and good mouse PK. Its activity against HIV isimproved as compared to 10E8V1.0/Pro140, but still slightly less thanthat of 10E8/Pro140. When paired with iMab in a CrossMab bispecific,this construct has double SEC peaks and poor mouse PK, and its activityagainst HIV is still roughly the same as 10E8/iMab and 10E8V1.0/iMab.10E8_(V2.0) is a chimeric antibody variant of 10E8 in which the FW1,CDR1 and part of FW2 are from 10E8_(V1.0) and in which the remainingpart of FW2, CDR2, FW3, CDR3 and FW4 are from 10E8. When paired withPro140 in a CrossMab bispecific, this construct has double SEC peaks andhas reduced activity against HIV as compared to 10E8/Pro140. When pairedwith iMab in a CrossMab bispecific, this construct has a single SECpeak, good PK, and activity against HIV that is improved over 10E8/iMab.10E8_(V3.0) is a somatic variant of 10E8 known as H11L1. H11L1/Pro140CrossMab has a single SEC peak and better anti-HIV activity than anyother 10E8/Pro140 construct (including the original one identified), buthas poor mouse PK due to autoreactivity. H11L1/iMab CrossMab has asingle SEC peak and anti-HIV activity that is better than the original10E8/iMab identified and roughly equivalent activity to that observedfor 10E8V2.0/iMab, but has poor mouse PK due to autoreactivity.

The variant of 10E8 that produced a single SEC peak in the context of aparticular CrossMab bispecific was different when paired with Pro140 oriMab. It appears that the stability of the 10E8 arm of these CrossMabbispecific antibodies is context dependent and will vary depending ofwhat antibody it is paired with. Thus, one variant (10E8_(V1.1)) wasidentified that was stable by SEC and with good mouse PK and goodanti-HIV activity when paired with Pro140. Another variant (10E8_(V2.0))was also identified that was stable by SEC with good mouse PK and withbetter anti-HIV activity than the originally identified 10E8/iMab.

Table 4 below describes the autoreactivity of tested variants, where“ANA” refers to anti-nuclear activity and “ACA” refers toanti-cardiolipin activity.

TABLE 4 Autoreactivity assessment in vitro Antibodies *Hep-2 (50 μg/mL)ANA ACA staining score Negative control − − Low positive control + +High positive control ++++ ++++ iMab − − ′ Pro140 − − ′ 10E8_(V1.0) − −0 10E8_(V1.1) − − ′ 10E8_(V2.0) − − ′ 10E8_(V3.0) −/+ +/++ 0.510E8_(V1.0)/P140 − − ′ 10E8_(V2.0)/Mab − − ′ 10E8_(V1.1)/P140 − − ′10E8_(V3.0)/iMab − − ′ 10E8_(V3.0)/P140 − −/+ ′

FIG. 30 depicts the pharmacokinetics of 10E8 and CrossMab antibodiesderived from several 10E8 variants and iMab or P140 in a mouse model. Asshown in FIGS. 31 and 32, 10E8_(v1.1)/P140 and 10E8_(v2.0)/iMab improveanti-HIV activity and stability, and have good stability when stored inPBS at 4° C. FIG. 33 depicts a native mass spectroscopy analysis of10E8_(v2.0)/iMab (N297A). FIG. 34 compares the activity of10E8_(v2.0)/P140 and 10E8_(v2.0)/iMab on a HIV Clade C panel, andcompares their IC50 and IC80 efficacy. FIGS. 35 and 36 compare thepotency of 10E8_(v1.1)/P140, 10E8_(v2.0)/iMab, and various monoclonalantibodies against HIV.

The terms and expressions employed herein are used as terms andexpressions of description and not of limitation, and there is nointention, in the use of such terms and expressions, of excluding anyequivalents of the features shown and described or portions thereof. Inaddition, having described certain embodiments of the invention, it willbe apparent to those of ordinary skill in the art that other embodimentsincorporating the concepts disclosed herein may be used withoutdeparting from the spirit and scope of the invention. Accordingly, thedescribed embodiments are to be considered in all respects as onlyillustrative and not restrictive.

1-20. (canceled)
 21. A method of treating HIV infection in a patient inneed thereof, comprising administering to the patient a pharmaceuticalcomposition comprising a bispecific antibody in a CrossMab formatcapable of neutralizing HIV, wherein the antibody comprises a lightchain and heavy chain portion of a first antibody 10E8, or a variantthereof, that binds to a HIV envelope protein, and a light chain andheavy chain portion of a second antibody ibalizumab, or a variantthereof, that binds to a cell membrane receptor protein or a cellmembrane co-receptor protein, wherein the light chain portion of thefirst antibody 10E8 comprises an amino acid sequence having at least 97%identity with SEQ ID NO: 33, and the heavy chain portion of the firstantibody 10E8 comprises an amino acid sequence having at least 97%identity with SEQ ID NO: 34; and the light chain portion of the secondantibody ibalizumab comprises an amino acid sequence having at least 97%identity with SEQ ID NO: 1, and the heavy chain portion of the secondantibody ibalizumab comprises an amino acid sequence having at least 97%identity with SEQ ID NO: 2; and wherein any amino acid alterationsrelative to SEQ ID NOS: 1, 2, 33, and 34 are outside of the variableregions.
 22. The method of claim 21, wherein the bispecific antibodycomprises a light chain portion of a 10E8 antibody comprising the aminoacid sequence of SEQ ID NO: 33 and a heavy chain portion of a 10E8antibody comprising the amino acid sequence of SEQ ID NO:
 34. 23. Themethod of claim 21, wherein the bispecific antibody comprises a lightchain portion of an ibalizumab antibody comprising the amino acidsequence of SEQ ID NO: 1 and a heavy chain portion of an ibalizumabantibody comprising the amino acid sequence of SEQ ID NO:
 2. 24. Amethod of treating HIV infection in a patient in need thereof,comprising administering to the patient a bispecific antibody in aCrossMab format capable of neutralizing HIV, wherein the antibodycomprises a light chain and heavy chain portion of a first antibody10E8, or a variant thereof, that binds to a HIV envelope protein, and alight chain and heavy chain portion of a second antibody Pro 140, or avariant thereof, that binds to a cell membrane receptor protein or acell membrane co-receptor protein, wherein the light chain portion ofthe first antibody 10E8 comprises an amino acid sequence having at least94% identity with SEQ ID NO: 31, and the heavy chain portion of thefirst antibody 10E8 comprises an amino acid sequence having at least 98%identity with SEQ ID NO: 32; and the light chain portion of the secondantibody Pro 140 comprises an amino acid sequence having at least 97%identity with SEQ ID NO: 11, and the heavy chain portion of the secondantibody Pro 140 comprises an amino acid sequence having at least 97%identity with SEQ ID NO: 12; and wherein any amino acid alterationsrelative to SEQ ID NOS: 11, 12, 31, and 32 are outside of the variableregions.
 25. The method of claim 24, wherein the bispecific antibodycomprises a light chain portion of a 10E8 antibody comprising the aminoacid sequence of SEQ ID NO: 31 and a heavy chain portion of a 10E8antibody comprising the amino acid sequence of SEQ ID NO:
 32. 26. Themethod of claim 24, wherein the bispecific antibody comprises a lightchain portions of a Pro 140 antibody comprising the amino acid sequenceof SEQ ID NO: 11 and a heavy chain portion of a Pro 140 antibodycomprising the amino acid sequence of SEQ ID NO: 12.